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Orchids produce millions of dust-like and reserveless tiny seeds per capsule and are unable to germinate on their own in nature. Orchid seeds have developed strategies to exploit seed germination-promoting orchid mycorrhizal fungi (sgOMFs) for acquisition of exogenous nutrients to fuel germination. Once colonized by sgOMFs, orchid seeds can digest intracellular sgOMF hyphae to acquire fungal nutrients, especially those for carbon nutrition. However, the underlying carbon delivery pathway from sgOMFs to orchid cells remains elusive. Clarification of the identities of fungally derived carbohydrates and the main molecular players responsible for the sgOMF-to-orchid carbon flow is of great significance for better understanding of symbiotic germination. To germinate in nature, orchid seeds strictly rely on seed germination-promoting orchid mycorrhizal fungi (sgOMFs) for provision of carbon nutrients. The underlying delivery pathway, however, remains elusive. We develop here a plausible model for sugar transport from sgOMFs to orchid embryonic cells to fuel germination. Orchids exploit sgOMFs to induce the formation of pelotons, elaborate intracellular hyphal coils in orchid embryos. The colonized orchid cells then obtain carbon nutrients by uptake from living hyphae and peloton lysis, primarily as glucose derived from fungal trehalose hydrolyzed by orchid-specific trehalases. The uptake of massive fungally derived glucose is likely to be mediated by two classes of membrane proteins, namely, sugars will eventually be exported transporters (SWEETs) and H+-hexose symporters. The proposed model serves as a launch pad for further research to better understand and improve orchid seed germination and conservation. [ABSTRACT FROM AUTHOR]

We carried out a circadian characterization of non-structural carbohydrate dynamics in the different organs of Erythrina velutina seedlings at late establishment. Seeds were incubated under controlled conditions for 9 days and then seedlings were hydroponically grown in a greenhouse for 8 days. Carbon dioxide uptake was measured in the cordiform leaves every 2 h during the daytime (12 h) and every 4 h during the night-time (12 h); seedlings were harvested every 4 h during the day-night cycle (24 h) to assess the contents of non-structural carbohydrates and the activities of amylases and invertases. Diel patterns of non-structural carbohydrate content were identified in the photosynthetic organs, in which the turnover of starch operated far from its depletion at dawn. In the heterotrophic organs, however, changes in the starch content over 24 h may have maintained the supply of soluble sugars, buffering transient fluctuations in carbon availability. Although it seems that amylase activity was not influenced by the circadian rhythm in the different seedling organs, diel patterns of invertase activity were recognized in the photosynthetic organs. [ABSTRACT FROM AUTHOR]

Background: Drought stress during flowering of maize (Zea mays L.) frequently results in decreased kernel setting, leading to grain yield depressions. Plasma membrane (PM) H+‐ATPase was identified as a key enzyme responsible for supply of assimilates to the developing maize kernels shortly after pollination. The activity of this enzyme was strongly inhibited under salt stress, pointing to an involvement in kernel abortion. Aims: This study aimed to determine whether also drought stress causes inhibition of PM H+‐ATPase in developing maize kernels shortly after pollination, leading to diminished hexose uptake and finally kernel abortion. The key questions are as follows: What are the limiting factors for grain yield production of maize plants facing drought? Are physiologically relevant parameters, quantified at flowering, reflected by yield determinants at maturity? Methods: Maize plants were cultivated using the container technique, and drought stress was imposed during 3 weeks bracketing flowering compared to well‐watered conditions throughout the entire growth period. The developing kernels were harvested 2 days after pollination, and PM vesicles were isolated and purified using two‐phase partitioning. Results: Water deficit caused a significant decrease in grain yield at maturity (−35%), which was determined by a reduced kernel number (−42%). Source limitation in the developing kernels under stress could be excluded. Acid invertase activity was unaffected by water deficit. Hexose availability was also no limiting factor for kernel setting and development. However, Vmax of in vitro hydrolytic activity of PM H+‐ATPase was significantly decreased in the developing maize kernels under drought stress and the maximal pH gradient at the PM was also significantly reduced. The observed inhibiting effects on PM H+‐ATPase were mainly of quantitative nature, as a lower number of proton pumps was present in the kernel PM. Qualitative changes of the enzyme (activation energy Ea, Michaelis constant Km) due to drought were not observed. Conclusions: The lower pH gradient probably decreased the proton‐driven transport of hexoses by carriers into the cytosol of the kernel cells, leading to kernel starvation and eventually contributing to kernel abortion. [ABSTRACT FROM AUTHOR]

Invertases are involved in plant growth, development, and stress adaptation; however, invertase-encoding genes have not yet been reported in Allium species. In this study, we identified 23 invertase izogenes in garlic (Allium sativum L.): 11 encoding putative neutral/alkaline (AsN/AINV1–11) and 12 acid (6 cell-wall—AsCWINV1–6 and 6 vacuolar—AsVINV1–6) enzymes. Among them, AsN/AINV1, 3, 8–10, AsCWINV2–5, and AsVINV2–6 showed significant transcription in garlic organs (roots, bulbs, pseudostems, leaves, sprouts, and reproductive parts) in a tissue-specific manner, whereas the AsN/AINV4–6, 11, AsCWINV1, 6, and AsVINV1 genes had weak or no detectable expression. Gene promoters contained nine, nine, and sixteen hormone-, stress-, and light-responsive cis-regulatory elements, respectively, and fifteen sites related to transcription factor binding and plant development. Expression analysis revealed that 12 invertase genes strongly transcribed in the roots of A. sativum cv. Ershuizao showed differential expression in the roots and leaves of A. sativum cv. Sarmat exposed to abiotic stresses (low temperature, high salinity, and drought) and phytohormones (abscisic acid and methyl jasmonate), which was significantly correlated with glucose, fructose, and sucrose contents. Our results should further functional analysis of invertases from Allium crops and contribute to the breeding of stress-tolerant varieties. [ABSTRACT FROM AUTHOR]

Background: Dendrobium officinale Kimura et Migo, a renowned traditional Chinese orchid herb esteemed for its significant horticultural and medicinal value, thrives in adverse habitats and contends with various abiotic or biotic stresses. Acid invertases (AINV) are widely considered enzymes involved in regulating sucrose metabolism and have been revealed to participate in plant responses to environmental stress. Although members of AINV gene family have been identified and characterized in multiple plant genomes, detailed information regarding this gene family and its expression patterns remains unknown in D. officinale, despite their significance in polysaccharide biosynthesis. Results: This study systematically analyzed the D. officinale genome and identified four DoAINV genes, which were classified into two subfamilies based on subcellular prediction and phylogenetic analysis. Comparison of gene structures and conserved motifs in DoAINV genes indicated a high-level conservation during their evolution history. The conserved amino acids and domains of DoAINV proteins were identified as pivotal for their functional roles. Additionally, cis-elements associated with responses to abiotic and biotic stress were found to be the most prevalent motif in all DoAINV genes, indicating their responsiveness to stress. Furthermore, bioinformatics analysis of transcriptome data, validated by quantitative real-time reverse transcription PCR (qRT-PCR), revealed distinct organ-specific expression patterns of DoAINV genes across various tissues and in response to abiotic stress. Examination of soluble sugar content and interaction networks provided insights into stress release and sucrose metabolism. Conclusions: DoAINV genes are implicated in various activities including growth and development, stress response, and polysaccharide biosynthesis. These findings provide valuable insights into the AINV gene amily of D. officinale and will aid in further elucidating the functions of DoAINV genes. [ABSTRACT FROM AUTHOR]

The ecotoxicity of Ag particles of different size has been assessed by microbiological, biochemical, and phytotoxic indicators for the upper layer (0–20 cm) of ordinary chernozem (Haplic Chernozem) in a laboratory model experiment. The effect has been studied of nano- (10 and 100 nm) and microparticles (1000 nm) of Ag at concentrations of 1, 10, and 100 mg/kg on the biological parameters of ordinary chernozem 30 days after contamination: the activity of catalase, dehydrogenases, ferrireductase, urease, peroxidase, polyphenol oxidase, invertase, phosphatase, the total number of bacteria, the abundance of bacteria of Azotobacter genus, the number of germinated seeds and the length of radish roots. It was found that the ecotoxicity of Ag particles depended on their size: in most cases, Ag particles 10 nm in size had a stronger ecotoxic effect on the biological parameters than particles 100 and 1000 nm in size. There were no significant differences in the ecotoxicity of 100 and 1000 nm Ag particles. The difference in the effects of Ag particles of different sizes increased with increasing Ag concentration in the soil: the higher the Ag concentration was in the soil (from 1 to 100 mg/kg), the more pronounced the difference was in ecotoxicity between 10 nm Ag particles and 100 and 1000 nm Ag particles. Phytotoxic indicators were more sensitive to contamination by Ag nanoparticles at all concentrations studied (1, 10 and 100 mg/kg); the total number of bacteria, invertase and phosphatase activity at 10 and 100 mg/kg; the abundance of bacteria of Azotobacter genus and the activity of dehydrogenases at 100 mg/kg. It is advisable to use these indicators in biodiagnostics of the ecotoxicity of Ag nanoparticles. [ABSTRACT FROM AUTHOR]

In this work, an acid invertase isolated from Lathyrus aphaca seedlings is purified, and its thermal properties are investigated. Acid invertase was purified through salt fractionation, CM-cellulose, and Ultrogel ACA-44 chromatography. Using SDS-PAGE, the consistency of the isolated enzyme was verified. The enzyme weighs 29 kDa at the molecular level. Acid invertase functions best at a pH of 3.0 and a temperature of 45 °C. Activation energy of 29.5 kJ mol−1 and K m and V max values of 0.5 mM and 119.7 µmol. min−1. mg−1 of protein was found for the isolated enzyme. It was shown that the mercaptide-forming agent p -chloromercuribenzoic acid (PCMB; 0.5 mM) and the enzyme's activity were both modestly increased by Ca2+, Mn2+, and Mg2+ ions but inhibited by Hg2+, Cd2+, and Pb2+ ions. At elevated temperatures, the invertase exhibited an increase in thermostability due to an increase in activation entropies (∆Sº) and a decrease in activation enthalpies (∆Hº). The interactions between 4 M urea and α-chymotrypsin were tetraphasic, causing periodic increases and decreases in invertase activity. It also suggests a potential explanation for acid-invertase thermal inactivation at high temperatures. This enzyme might be a catabolite-resistant invertase to produce high-gravity ethanol or fructose syrup. [Display omitted] • Acid invertase was purified from Lathyrus aphaca seedlings with molecular weight 29 kDa. • Acid invertase free in a solution of 5% (w/v) sucrose, pH 3.0, is stable up to 50 °C for a period of 4 h. • The K m and V max values for acid invertase was 0.5 mM and 119.7 µmol. min-1.mg−1 of protein. • The acid invertase is more stable and their energy of deactivation are 107 kJ/mol (25 kcal/mol), respectively. • The effects of α-chymotrypsin and 4 M urea were tetraphasic with periodic gain and loss of enzyme activity. [ABSTRACT FROM AUTHOR]

Potato (Solanum tuberosum) is the third most important food crop in the world. Potato tubers must be stored at cold temperatures to minimize sprouting and losses due to disease. However, cold temperatures strongly induce the expression of the potato vacuolar invertase gene (VInv) and cause reducing sugar accumulation. This process, referred to as "cold-induced sweetening," is a major postharvest problem for the potato industry. We discovered that the cold-induced expression of VInv is controlled by a 200 bp enhancer, VInvIn2En, located in its second intron. We identified several DNA motifs in VInvIn2En that bind transcription factors involved in the plant cold stress response. Mutation of these DNA motifs abolished VInvIn2En function as a transcriptional enhancer. We developed VInvIn2En deletion lines in both diploid and tetraploid potato using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9)-mediated gene editing. VInv transcription in cold-stored tubers was significantly reduced in the deletion lines. Interestingly, the VInvIn2En sequence is highly conserved among distantly related Solanum species, including tomato (Solanum lycopersicum) and other non-tuber-bearing species. We conclude that the VInv gene and the VInvIn2En enhancer have adopted distinct roles in the cold stress response in tubers of tuber-bearing Solanum species. An intronic enhancer is responsible for cold-induced expression of the vacuolar invertase gene in potato. [ABSTRACT FROM AUTHOR]

Sugar beet molasses is a low-value byproduct from the sugar industry. It contains significant amounts of sucrose (approx. 50% (w/w)), which can be used for many different applications, for example, as feedstock for the production of fuel (as ethanol) and biobased chemicals such as 5-hydoxymethyl furfural (HMF). To produce platform chemicals, sucrose is hydrolyzed into its monomeric C6 sugars: glucose and fructose. When comparing the hydrolysis rates of molasses with a pure sucrose solution, the specific reaction rate is much slower (Qp/x,60min = 93 and 70 gprod L−1 h−1 gcell−1 for pure sucrose and crude molasses, respectively) at the same sucrose concentration (300 g/L) and process conditions. To clarify why molasses inhibits the enzymatic hydrolysis rate, the influence of its viscosity and inorganic and organic composition was investigated. Also, the effects of molasses and treated molasses on pure enzymes, invertase (from Saccharomyces cerevisiae, 0.05 mg/mL), compared with hydrolysis using whole cells of Baker's yeast (3 mg/mL), were tested. The results indicate an inhibitory effect of potassium (Qp/x,60min = 76 gprod L−1 h−1 gcell−1), generally at high salt concentrations (Qp/x,60min = 67 gprod L−1 h−1 gcell−1), which could be correlated to the solution's high salt concentrations and possibly the synergistic effects of different ions when applying concentrations that were four times that in the molasses. Also, the viscosity and sucrose purity seem to have an effect, where pure sucrose solutions and thick juice from the sugar mill yielded higher hydrolysis rates (Qp/x,60min = 97 gprod L−1 h−1 gcell−1) than molasses-type solutions with a higher viscosity (Qp/x,60min = 70–74 gprod L−1 h−1 gcell−1). Attempting to further understand the effects of different components on the invertase activity, an in silico investigation was performed, indicating that high salt concentrations affected the binding of sucrose to the active site of the enzyme, which can result in a lower reaction rate. This knowledge is important for future scale-up of the hydrolysis process, since reduced hydrolysis rates require larger volumes to provide a certain productivity, requiring larger process equipment and thereby higher investment costs. [ABSTRACT FROM AUTHOR]

As one of the biomarkers of coagulation system-related diseases, the detection of thrombin is of practical importance. Thus, this study developed a portable biosensor based on a personal glucometer for rapid detection of thrombin activity. Fibrinogen was used for the detection of thrombin, and the assay principle was inspired by the blood coagulation process, where thrombin hydrolyzes fibrinogen to produce a fibrin hydrogel, and the amount of invertase encapsulated in the fibrin hydrogel fluctuates in accordance with the activity of thrombin in the sample solution. The quantitative assay is conducted by measuring the amount of unencapsulated invertase available to hydrolyze the substrate sucrose, and the signal readout is recorded using a personal glucometer. A linear detection range of 0–0.8 U/mL of thrombin with a limit of detection of 0.04 U/mL was obtained based on the personal glucometer sensing platform. The results of the selectivity and interference experiments showed that the developed personal glucometer sensing platform is highly selective and accurate for thrombin activity. Finally, the reliability of the portable glucometer method for rapid thrombin detection in serum samples was investigated by measuring the recovery rate, which ranged from 92.8% to 107.7%. In summary, the fibrin hydrogel sensing platform proposed in this study offers a portable and versatile means for detecting thrombin using a personal glucometer. This approach not only simplifies the detection process, but also eliminates the need for large instruments and skilled operators, and substantially reduces detection costs. [ABSTRACT FROM AUTHOR]

Bamboo shoots are considered as a healthy food and are popular in Asian cooking. The distribution of nutrients and their dynamics during the growth of bamboo shoots play a crucial role in guiding the harvesting and processing of bamboo shoots. In this study, Dendrocalamus brandisii (Munro) Kurz, an important bamboo species for harvesting fine edible shoots in southern China and Southeast Asia, was used to measure several indicators related to the edibility and nutritional value of fresh shoots across five height grades (H1: 20~30 cm; H2: 31~40 cm; H3: 41~50 cm; H4: 51~60 cm; and H5: 61~70 cm). The results indicated that, although the nutrient and mineral elements showed an increasing (crude fiber content, CFC), decreasing (total soluble sugars content, TSSC; ash content, AC; Fe; Zn), or fluctuating (soluble protein content, SPC; P; K; Ca) pattern with the growth of bamboo shoots, both the CFC and TSSC showed the highest values in the lower part at five growth heights, indicating that carbohydrates were mainly enriched in the bases of bamboo shoots. The SPC, AC, and other mineral elements were higher in the upper part, especially in H1–H3. Combined with the relatively high activity of metabolic enzymes (sucrose-phosphate synthase, SPS; neutral invertase, NI) in the upper part of bamboo shoots (although not statistically significant), it can be inferred that the shoot tip may be the main tissue for early nutrient synthesis and metabolism. Compared with the shoot height, different parts of a D. brandisii bamboo shoot are more indicative of its nutrient properties. Although all heights of bamboo shoots showed an abundance of nutrients and mineral elements, bamboo shoots with a height of less than 40 cm had a higher TSSC, AC, Fe, and Zn, and a lower CFC, thus having a better balance between nutrients and edibility. [ABSTRACT FROM AUTHOR]

Abstract Background Dendrobium officinale Kimura et Migo, a renowned traditional Chinese orchid herb esteemed for its significant horticultural and medicinal value, thrives in adverse habitats and contends with various abiotic or biotic stresses. Acid invertases (AINV) are widely considered enzymes involved in regulating sucrose metabolism and have been revealed to participate in plant responses to environmental stress. Although members of AINV gene family have been identified and characterized in multiple plant genomes, detailed information regarding this gene family and its expression patterns remains unknown in D. officinale, despite their significance in polysaccharide biosynthesis. Results This study systematically analyzed the D. officinale genome and identified four DoAINV genes, which were classified into two subfamilies based on subcellular prediction and phylogenetic analysis. Comparison of gene structures and conserved motifs in DoAINV genes indicated a high-level conservation during their evolution history. The conserved amino acids and domains of DoAINV proteins were identified as pivotal for their functional roles. Additionally, cis-elements associated with responses to abiotic and biotic stress were found to be the most prevalent motif in all DoAINV genes, indicating their responsiveness to stress. Furthermore, bioinformatics analysis of transcriptome data, validated by quantitative real-time reverse transcription PCR (qRT-PCR), revealed distinct organ-specific expression patterns of DoAINV genes across various tissues and in response to abiotic stress. Examination of soluble sugar content and interaction networks provided insights into stress release and sucrose metabolism. Conclusions DoAINV genes are implicated in various activities including growth and development, stress response, and polysaccharide biosynthesis. These findings provide valuable insights into the AINV gene amily of D. officinale and will aid in further elucidating the functions of DoAINV genes.

Background and Aims Extrafloral nectaries are nectar-secreting structures present on vegetative parts of plants which provide indirect defences against herbivore attack. Extrafloral nectaries in Clerodendrum chinense are patelliform-shaped specialized trichomatous structures. However, a complete understanding of patelliform extrafloral nectaries in general, and of C. chinense in particular, has not yet been established to provide fundamental insight into the cellular physiological machinery involved in nectar biosynthesis and secretory processes. Methods We studied temporal changes in the morphological, anatomical and ultrastructural features in the architectures of extrafloral nectaries. We also compared metabolite profiles of extrafloral nectar, nectary tissue, non-nectary tissue and phloem sap. Further, both in situ histolocalization and normal in vitro activities of enzymes related to sugar metabolism were examined. Key Results Four distinct tissue regions in the nectar gland were revealed from histochemical characterization, among which the middle nectariferous tissue was found to be the metabolically active region, while the intermediate layer was found to be lipid-rich. Ultrastructural study showed the presence of a large number of mitochondria along with starch-bearing chloroplasts in the nectariferous region. However, starch depletion was noted with progressive maturation of nectaries. Metabolite analysis revealed compositional differences among nectar, phloem sap, nectary and non-nectary tissue. Invertase activity was higher in secretory stages and localized in nectariferous tissue and adjacent region. Conclusions Our study suggests extrafloral nectar secretion in C. chinense to be both eccrine and merocrine in nature. A distinct intermediate lipid-rich layer that separates the epidermis from nectary parenchyma was revealed, which possibly acts as a barrier to water flow in nectar. This study also revealed a distinction between nectar and phloem sap, and starch could act as a nectar precursor, as evidenced from enzymatic and ultrastructural studies. Thus, our findings on changing architecture of extrafloral nectaries with temporal secretion revealed a cell physiological process involved in nectar biosynthesis and secretion. [ABSTRACT FROM AUTHOR]

This study investigated the production of bakery and sweet paste products containing the prebiotic fructooligosaccharide (FOS) using an in situ method with invertase. The FOS formation method was optimized for each product to ensure high quality and appropriate sweetness. The method effectively decreased the sugar content in the final product by 12.7–68.4% while maintaining quality. The FOS content was 3.8–4.8% in castella, 0.6–3.6% in sweet dough bread, and 7.5–8.5% in sweet chickpea paste. By contrast, the commercial method of adding FOS decreased product quality; castella product height decreased by 20.8%, and hardness increased by 79%. The specific volume of the sweet dough bread decreased by 17.4% and hardness increased by 59%. Therefore, we developed a commercially feasible method to efficiently utilize FOS in sugar-containing foods while maintaining their quality. [ABSTRACT FROM AUTHOR]

Strigolactones (SLs) were recently defined as a novel class of plant hormones that act as key regulators of diverse developmental processes and environmental responses. Much research has focused on SL biosynthesis and signaling in roots and shoots, but little is known about whether SLs are produced in early developing seeds and about their roles in ovule development after fertilization. This study revealed that the fertilized ovules and early developing pericarp in Xanthoceras sorbifolium produced minute amounts of two strigolactones: 5-deoxystrigol and strigol. Their content decreased in the plants with the addition of exogenous phosphate (Pi) compared to those without the Pi treatment. The exogenous application of an SL analog (GR24) and a specific inhibitor of SL biosynthesis (TIS108) affected early seed development and fruit set. In the Xanthoceras genome, we identified 69 potential homologs of genes involved in SL biological synthesis and signaling. Using RNA-seq to characterize the expression of these genes in the fertilized ovules, 37 genes were found to express differently in the fertilized ovules that were aborting compared to the normally developing ovules. A transcriptome analysis also revealed that in normally developing ovules after fertilization, 12 potential invertase genes were actively expressed. Hexoses (glucose and fructose) accumulated at high concentrations in normally developing ovules during syncytial endosperm development. In contrast, a low ratio of hexose and sucrose levels was detected in aborting ovules with a high strigolactone content. XsD14 virus-induced gene silencing (VIGS) increased the hexose content in fertilized ovules and induced the proliferation of endosperm free nuclei, thereby promoting early seed development and fruit set. We propose that the crosstalk between sugar and strigolactone signals may be an important part of a system that accurately regulates the abortion of ovules after fertilization. This study is useful for understanding the mechanisms underlying ovule abortion, which will serve as a guide for genetic or chemical approaches to promote seed yield in Xanthoceras. [ABSTRACT FROM AUTHOR]

Enzymatic activity of soils is the most important diagnostic indicator of the ecological state of soils affected by various types of anthropogenic impact. The aim of the study was to evaluate the enzymatic activity of ordinary chernozem (Haplic Chernozem) contaminated with Ag, Bi, Te, and Tl. Ten enzymes (catalase, dehydrogenase, peroxidase, polyphenol oxidase, ascorbate oxidase, ferrireductase, protease, phosphatase, invertase, and urease) were analyzed. According to the degree of inhibition of enzymes, heavy metals formed the following sequence: Tl > Ag > Bi > Te. With an increase in the concentration of heavy metals, the toxic effect on the activity of enzymes increased. The oxidoreductases showed greater sensitivity to Ag, Bi, Te, and Tl contamination than hydrolases. Among oxidoreductases, the highest sensitivity was found for ferrireductase, and the lowest one for ascorbate oxidase. According to the activity of enzymes of the hydrolase class, invertase was the most sensitive, and urease was the least sensitive. When contaminated with Ag, Bi, and Te, invertase had the highest informative value, and when contaminated with Tl, urease and polyphenol oxidase were the most informative. Among the enzymes of the oxidoreductase class, the highest information value was found for peroxidase, and the lowest one for ascorbate oxidase. Among the enzymes of the hydrolase class, invertase was the most sensitive, and phosphatase was the least sensitive. The results of the study can be used to assess the ecological state of soils contaminated with Ag, Bi, Te and Tl. [ABSTRACT FROM AUTHOR]

Sugarcane is a significant primitive source of sugar and energy worldwide. The progress in enhancing the sugar content in sugarcane cultivars remains limited due to an insufficient understanding of specific genes related to sucrose production. The present investigation examined the enzyme activities, levels of reducing and non-reducing sugars, and transcript expression using RT-qPCR to assess the gene expression associated with sucrose metabolism in a high-sucrose sugarcane clone (GXB9) in comparison to a low-sucrose sister clone (B9). Sucrose phosphate synthase (SPS), sucrose phosphate phosphatase (SPP), sucrose synthase (SuSy), cell wall invertase (CWI), soluble acid invertase (SAI), and neutral invertase (NI) are essential enzymes involved in sucrose metabolism in sugarcane. The activities of these enzymes were comparatively quantified and analyzed in immature and maturing internodes of the high- and low-sucrose clones. The results showed that the higher-sucrose-accumulating clone had greater sucrose concentrations than the low-sucrose-accumulating clone; however, maturing internodes had higher sucrose levels than immature internodes in both clones. Hexose concentrations were higher in immature internodes than in maturing internodes for both clones. The SPS and SPP enzymes activities were higher in the high-sucrose-storing clone than in the low-sucrose clone. SuSy activity was higher in the low-sucrose clone than in the high-sucrose clone; further, the degree of SuSy activity was higher in immature internodes than in maturing internodes for both clones. The SPS gene expression was considerably higher in mature internodes of the high-sucrose clones than the low-sucrose clone. Conversely, the SuSy gene exhibited up-regulated expression in the low-sucrose clone. The enhanced expression of SPS in the high-sucrose clone compared to the low-sucrose clone suggests that SPS plays a major role in the increased accumulation of sucrose. These findings provide the opportunity to improve sugarcane cultivars by regulating the activity of genes related to sucrose metabolism using transgenic techniques. [ABSTRACT FROM AUTHOR]

The speedy growth of human population, the issues arising out of pollution, demand of invertase yield and dumping of wastes from agro-industry have been influenced to find new solutions for microbial enzymes. In this study, an attempt was made to produce the invertase by yeast isolated from honey bees. Influence of various process parameters like nutritional sources and environmental conditions was studied using waste material. The enzyme was precipitated by using the acetone to get the product for further experimental analysis. Subsequently, it was carried out for further purification by Sephadex G-50 column chromatography and dialysed. At that moment, optimization of culture conditions for the yield of invertase with agro waste was determined by process parameters. It was followed by the immobilization of microbial cells for increasing the yield of invertase by various matrices. In order to check the bioactive potential of invertase, the bread was prepared with and without the amendment of enzyme. The existing result could be supportive to future researchers for industrial applications of microbial invertase by immobilization approach. [ABSTRACT FROM AUTHOR]

The results of studying the effect of fires on biological properties of cinnamonic soils of xerophytic forests (Skeletic Leptic Cambisol) of the Utrish State Nature Reserve, Krasnodar krai (crown fire of 2020), and burozem of mesophytic forests (Haplic Cambisol (Loamic)) of the Khamyshinsk forestry, Republic of Adygea (ground fire of 2018) are presented. Changes in the soil reaction, in the organic carbon content, and in the activity of enzymes: catalase, urease, phosphatase, and invertase involved in the cycle of carbon, phosphorus, and nitrogen have been studied. The reaction of enzymes to pyrogenic effects depends on the enzyme kind and the soil type. The factor analysis was also performed. The activity of catalase and invertase decreases by 47% on average for two soil types in the 0–3-cm-thick layer, while the reaction of phosphatase and urease depends on the soil type. Two years after the fire, the phosphatase activity in the surface layer of post-pyrogenic cinnamonic soils approached the control values; and the urease activity regenerated more slowly as compared to other enzymes. Four years after the fire, the values of urease activity in the 0–3-cm-thick layer of the post-pyrogenic acid burozem, on the contrary, approached the control values. An increase in pH by 30% on average and a decrease in the Corg content by 12% on average were also recorded for both two soil types. The mean activity of all the studied enzymes in the 3–10-cm-thick layer increased in cinnamonic soils and, on the contrary, decreased in acid burozem. The factor analysis showed a relationship between the soil reaction, the organic carbon content, and the activity of enzymes, and the strength and nature of relationships differed depending on the soil type. The results obtained testify to the effect of soil properties on the response of enzymatic activity to the pyrogenic impact. [ABSTRACT FROM AUTHOR]

Carbohydrates and gamma-amino butyric acid (GABA) have not been studied under the stress of stripe rust caused by Puccinia striiformis f. sp. tritici (Pst) in wheat. Wheat cultivars PBW725 (resistant) and HD2967 (susceptible) were compared at 1, 2 and 4 d of GABA, Pst, and GABA + Pst sprayed to 30-d of seedling growth. Starch was hydrolyzed, and levels of sugars and proteins improved over time of infection in PBW725 however HD2967 improved under GABA + Pst. Proline increased while GABA decreased over the time of infection in HD2967. Progressive increase of diamine oxidase and glutamate decarboxylase was seen in PBW725 during infection. Protease levels became low at 4 d of infection but improved under GABA + Pst in HD2967. Protein carbonyls increased in HD2967 only under Pst but decreased under GABA + Pst. Results concluded that GABA is an essential regulator of carbohydrates under stripe rust. Sustained response over time of infection may be important for resistance. [ABSTRACT FROM AUTHOR]

Key message: Sucrose invertase activity is positively related to osmotic and salt stress resistance in peanut. Sucrose invertases (INVs) have important functions in plant growth and response to environmental stresses. However, their biological roles in peanut are still not fully revealed. In this research, we identified 42 AhINV genes in the peanut genome. They were highly conserved and clustered into three groups with 24 segmental duplication events occurred under purifying selection. Transcriptional expression analysis exhibited that they were all ubiquitously expressed, and most of them were up-regulated by osmotic and salt stresses, with AhINV09, AhINV23 and AhINV19 showed the most significant up-regulation. Further physiochemical analysis showed that the resistance of peanut to osmotic and salt stress was positively related to the high sugar content and sucrose invertase activity. Our results provided fundamental information on the structure and evolutionary relationship of INV gene family in peanut and gave theoretical guideline for further functional study of AhINV genes in response to abiotic stress. [ABSTRACT FROM AUTHOR]

The food enzyme β‐fructofuranosidase (β‐d‐fructofuranoside fructohydrolase; EC 3.2.1.26) is produced with the non‐genetically modified Saccharomyces cerevisiae strain NCYC R693 by Kerry Ingredients & Flavours Ltd. The production strain meets the requirements for the qualified presumption of safety (QPS) approach. The food enzyme is intended to be used in four food manufacturing processes. The dietary exposure to the food enzyme–total organic solids (TOS) was estimated to be up to 2.485 mg TOS/kg body weight per day in European populations. As the production strain qualifies for the QPS approach of safety assessment and no issue of concern arising from the production process of the food enzyme were identified, the Panel considered that no toxicological studies other than the assessment of allergenicity were necessary. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and one match with a tomato allergen was found. The Panel considered that the risk of allergic reactions upon dietary exposure to this food enzyme, particularly in individuals sensitised to tomato, cannot be excluded. However, the likelihood of allergic reactions is expected not to exceed the likelihood of allergic reactions to tomato. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use. [ABSTRACT FROM AUTHOR]

Biomimetic ligands are synthetic compounds that mimic the structure and binding properties of natural biological ligands. The first uses of textile dyes as pseudo-affinity ligands paved the way for the rational design and de novo synthesis of low-cost, non-toxic and highly stable triazine-scaffolded affinity ligands. A novel method to assess and enhance protein stability, employing triazine-based biomimetic ligands and using cutinase from Fusarium solani pisi as a protein model, has been previously reported. This innovative approach combined the concepts of molecular modeling and solid-phase combinatorial chemistry to design, synthesize and screen biomimetic compounds able to bind cutinase through complementary affinity-like interactions while maintaining its biological functionality. The screening of a 36-member biased combinatorial library enabled the identification of promising lead ligands. The immobilization/adsorption of cutinase onto a particular lead (ligand 3′/11) led to a noteworthy enhancement in thermal stability within the temperature range of 60–80 °C. In the present study, similar triazine-based compounds, sourced from the same combinatorial library and mimicking dipeptides of diverse amino acids, were selected and studied to determine their effectiveness in binding and/or improving the thermal stability of several lipases, enzymes which are closely related in function to cutinases. Three ligands with different compositions were screened for their potential thermostabilizing effect on different lipolytic enzymes at 60 °C. An entirely distinct enzyme, invertase from Saccharomyces cerevisiae, was also assessed for binding to the same ligands and functioned as a 'control' for the experiments with lipases. The high binding yield of ligand 3′/11 [4-({4-chloro-6-[(2-methylbutyl)amino]-1,3,5-triazin-2-yl}amino)benzoic acid] to cutinase was confirmed, and the same ligand was tested for its ability to bind lipases from Aspergillus oryzae (AOL), Candida rugosa (CRL), Chromobacterium viscosum (CVL), Rhizomucor miehei (RML) and Rhizopus niveus (RNL). The enzymes CRL, CVL, RNL and invertase showed significant adsorption yields to ligand 3′/11—32, 29, 36 and 94%, respectively, and the thermal stability at 60 °C of free and adsorbed enzymes was studied. CVL and RNL were also stabilized by adsorption to ligand 3′/11. In the case of CRL and invertase, which bound but were not stabilized by ligand (3′/11), other ligands from the original combinatorial library were tested. Between the two alternative ligands, one was effective at stabilizing C. rugosa lipase, while none stabilized invertase. [ABSTRACT FROM AUTHOR]

This article discusses two studies that explore methods for reducing acrylamide levels in processed potatoes. The first study used New Genomic Techniques (NGTs) to edit the genetic makeup of a potato variety called Lady Rosetta, resulting in lower levels of acrylamide. The second study focused on identifying specific genetic variations in potato alleles that can reduce acrylamide content in processed products. Both studies offer potential solutions for reducing acrylamide without the use of harmful chemicals. The data supporting these findings can be found in the supplementary material of the article. [Extracted from the article]

With the objective of understanding microbe—pesticide interactions in soil, the effect of pesticides alone and in combination on the activities of amylase, cellulase, and invertase was assessed in black, and red soils collected from the groundnut (Arachis hypogaea L.) cultivated fields of Andhra Pradesh, India. In addition, the potential bacteria that can break down chlorpyrifos was identified using biochemical assays and a partial 16S rRNA gene sequence analysis. Soil enzymes were analyzed using standard procedures described elsewhere. The activities of amylase, cellulase, and invertase were solely dependent on the dose of pesticides used in the current study. Enzyme activities were significantly increased at the level of 2.5 or 5.0 kg ha–1, whereas increases in the concentration of pesticides at 7.5–10 kg ha−1 were lethal to the enzyme activities in both soils at 10-day incubation. The effective combination for the increase in the activities of amylase, cellulase, and invertase were monocrotophos + mancozeb. Mineral salts medium with 50 ppm chlorpyrifos as a carbon source had the greatest growth of the bacteria (i.e. Pseudomonas citronellolis strain CF3) at 14th h of incubation with OD value of 0.24 at 660 nm, and the same strain could eliminate 75% of the chlorpyrifos within 24 h from the medium. Based on the present study it was concluded that insecticides used alone or in combination with fungicides at specific doses (2.5 or 5.0 kg ha−1) improved the amylase, cellulase, and invertase activities, which are involved in the carbon cycle in soils. Pseudomonas citronellolis strain CF3 is a potential bacterium in the remediation of chlorpyrifos-contaminated sites.

Sugar beet factories employ long-term storage of sugar beet roots to extend the processing campaign in Egypt. Changes in the chemical composition of sugar beet roots during storage, driven by invertase enzyme activity, impact the crystallization process in sugar production. This study aimed to monitor invertase activity in sugar beet roots harvested at 180, 195 and, 210 days of cultivation, applying different storage conditions for 1, 10, 20, and 30 days. Additionally, the study explored the influence of invertase activity on reducing sugars content in sugar beet roots. Results indicated that covered storage conditions exhibited the lowest invertase activity compared to open air and room storage conditions. Moreover, reducing sugars content was affected by invertase enzyme activity. Storage conditions displayed the lowest inverted sugars content across all tested sugar beet root varieties. Thus, the study suggests that long-term storage with covered conditions is suitable for extending the processing campaign, emphasizing the importance of considering inverted sugars concentration. [ABSTRACT FROM AUTHOR]

Invertase is a hydrolytic enzyme that causes the glyosidic connection between glucose and fructose in sucrose to break, forming an invert sugar that rotates polarized light from the right to the left. Because of its unbound features, invertase has become an important industrial enzyme. A total of four bacterial isolates were isolated from deteriorating apple and banana peel collected from nearby fruit market vendors. Invertase production was investigated through screening and biochemical analysis. As a result, it was discovered that A1 and B1 isolates produced sufficient invertase for industrial use. For selecting a range of parameters to fit in Central composite Design and analysis to be done using Response surface Methodology, the One Variable at a Time approach was chosen. Elite EkINV and BsINV production were observed at 25 and 45 °C incubation temperature in the media comprised of variable concentration of carbon source, nitrogen source and inoculated culture according to the experimental and statistical findings. The experiment's coefficient of determination (R2) values of 0.73 and 0.82 indistinctively resulted in a good fit of the model. Isolate A1 was identified as Enterobcter kobei and isolate B1 as Bacillus Subtilis Strain with accession numbers MN267695 and MN263256, respectively, after 16s rRNA molecular sequencing. [ABSTRACT FROM AUTHOR]

The application of wounding stress can induce the accumulation of phenolic antioxidants in carrots. This study aimed to investigate the possible regulation role of invertase (INV) on the biosynthesis of phenolics in wounded carrots. In this study, carrots were cut into two different wounding intensities of slices and cubes, then stored at 20 °C for 2 days. The results showed that wounding stress caused an obvious increase in phenolic content and antioxidant capacity in carrot tissues, and a positive correlation was observed between the biosynthesis of phenolic compounds and the degradation of sucrose. Simultaneously, wounding activated the sucrose-cleaving enzymes of INVs, including acid INV (AI) and neutral INV (NI), and up-regulated the expressions of most encoding genes of INVs. In addition, treatment with INV activators accelerated the accumulation of phenolic antioxidants, while treatment with INV inhibitors suppressed this process, suggesting that the synthesis of phenolic compounds in wounded carrots is closely related to the availability of sugars. Our findings provide new insights into the regulation role of INV on the wound-induced accumulation of phenolic compounds in carrots, which may be helpful in using wounded plants to produce more phenolic antioxidants. [ABSTRACT FROM AUTHOR]

AbstractEkINV and BsINV are two invertases from Enterobacter kobei and Bacillus subtillis, respectively. The two enzymes were produced extracellularly at pH 3.0 and former at 45 °C while latter at 25 °C. The Km values were observed to be 1.7 mol/L (EkINV) and 1.9 mol/L (BsINV). Purification and characterization analysis revealed trimer structure EkINV having subunit molecular mass as 87.14, 42.98 and 29.0 kDa. Unlike, BsINV was monomer with molecular mass of 55.1 kDa. The invertases beside hydrolytic activity exhibited transfructolysation which is used in preparation of low calorie Ready To Serve mango juice and muffins. [ABSTRACT FROM AUTHOR]

Exogenous nitrogen and carbon can affect plant cell walls, which are composed of structural carbon. Sucrose synthase (SUS), invertase (INV), hexokinase (HXK), phosphoglucomutase (PGM), and UDP-glucose pyrophosphorylase (UGP) are the key enzymes of sucrose metabolism involved in cell wall synthesis. To understand whether these genes are regulated by carbon and nitrogen to participate in structural carbon biosynthesis, we performed genome-wide identification, analyzed their expression patterns under different carbon and nitrogen treatments, and conducted preliminary functional verification. Different concentrations of nitrogen and carbon were applied to poplar (Populus trichocarpa Torr. and Gray), which caused changes in cellulose, lignin, and hemicellulose contents. In poplar, 6 SUSs, 20 INVs, 6 HXKs, 4 PGMs, and 2 UGPs were identified. Moreover, the physicochemical properties, collinearity, and tissue specificity were analyzed. The correlation analysis showed that the expression levels of PtrSUS3/5, PtrNINV1/2/3/5/12, PtrCWINV3, PtrVINV2, PtrHXK5/6, PtrPGM1/2, and PtrUGP1 were positively correlated with the cellulose content. Meanwhile, the knockout of PtrNINV12 significantly reduced the cellulose content. This study could lay the foundation for revealing the functions of SUSs, INVs, HXKs, PGMs, and UGPs, which affected structural carbon synthesis regulated by nitrogen and carbon, proving that PtrNINV12 is involved in cell wall synthesis. [ABSTRACT FROM AUTHOR]

The enzymatic approach is a highly effective and the major scientific method to eliminating bitter components in citrus-derived products nowadays. Microbial production of limonin invertase stands out due to its pivotal role in the removal of the bitter substance, limonin. The optimization of fermentation parameters and the study of scale-up fermentation are imperative for product commercialization. In this study, we focused on optimizing stirring speed, fermentation temperature, and initial pH to enhance the growth and limonin invertase production by the Aspergillus tabin strain UA13 in a 5-l stirred-tank bioreactor. Our results revealed the following optimal parameters are: a stirring speed of 300 rpm, a fermentation temperature of 35°C and a pH 5.0. Under these optimized conditions, the limonin invertase activity reached its peak at 63.38 U ml−1, representing a 1.67-fold increase compared to the unoptimized conditions (38.10 U ml−1), while also reducing the fermentation duration by 12 h. Furthermore, our research demonstrated that limonin invertase effectively hydrolyze limonin in grapefruit juice, reducing its content from 13.28 to 2.14 μg ml−1, as determined by HPLC, resulting in a 6.21-fold reduction of the bitter substance. [ABSTRACT FROM AUTHOR]

Sugarcane (Saccharum spp. hybrid) is the chief source of sugar and biofuel globally and is prominent among cash crops. Sucrose is the main required product in sugarcane, and many studies have been performed to understand the phenomena of sucrose synthesis, metabolism, and accumulation in sugarcane. However, none of the studies concluded that a single gene is responsible for the sucrose content. Instead, a complex mechanism consisting of several genes, such as sucrose phosphate synthase genes (SPS1, SPS2, SPS4, SPS5), sucrose synthase genes (SuSy1, SuSy2, SuSy4), invertase genes (INV, CWIN, NIN1, CINV2), and phytohormone, trehalose, transcription factor (TF), protein kinase, and sugar transporter genes are working spatiotemporally in sugarcane. Currently, omics approaches like transcriptomics, proteomics, and metabolomics are also being used to explore the sugar metabolism in sugarcane, but integrated transcriptomic, proteomic, and metabolomic studies have been less reported. The results obtained from the integrated analysis of transcriptomics, proteomics, and metabolomics are more reliable because the strong gene expression, received in the form of abundant mRNA, does not guarantee the plentiful existence of associated proteins or their particular activity in the target cells or tissues, which discloses the restraint of single interpretation and stresses the significance of the integrated analysis of transcriptomics, proteomics, and metabolomics. This review highlights different genes and molecules contributing to sugar metabolism at different stages and the significance of omics approaches in explaining sucrose metabolism, especially sucrose accumulation in sugarcane. It is also a vital source of knowledge for sugarcane breeders, particularly associated with sucrose content improvement and bioethanol energy production. [ABSTRACT FROM AUTHOR]

The article presents the results of studies on the species composition of vascular plants and soil enzymatic activity in communities dominated by Acer negundo. The studies were carried out on accounting sites located in the mountain-taiga and forest-steppe zones of Kuzbass. The selection of trial plots was made taking into account the horizontal differentiation of space into under-crown (registration plots) and external zones (control) of phytogenic fields. Floristic descriptions and the level of activity of enzymes (invertase, protease, and phosphatase) in the soil were analyzed. The diversity of vascular plants in the areas under study constituted 64 species. The dominant plant species in the ash-leaved maple communities were identified. Ruderal species Cirsium setosum, Elytrigia repens, Lamium album, Urtica dioica, and Taraxacum officinale dominate under the canopy of A. negundo, while the proportion of meadow species Achillea millefolium, Dactylis glomerata, and Festuca pratensis increases in the outer zones. It has been shown that the floristic composition of communities does not affect significantly the level of activity of soil enzymes. An increase in the activity of hydrolytic enzymes by an average of 11% compared with the outer zones was noted in the under-crown space of A. negundo. [ABSTRACT FROM AUTHOR]

In this work, we established an efficient process for the production of itaconate from the regionally sourced industrial side-stream molasses using Ustilago cynodontis and Ustilago maydis. While being relatively cheap and more environmentally friendly than refined sugars, there are some major challenges to overcome when working with molasses. Some of those challenges are a high nitrogen load, unknown impurities in the feedstock, and high amounts of ill-favoured carbon sources, such as sucrose or lactate. We could show that the activity of the sucrose-hydrolysing enzyme invertase plays a crucial role in the efficiency of the process and that the fructose utilisation differs between the two strains used in this work. Thus, with a higher invertase activity, the ability to convert fructose into the desired product itaconate, and an overall higher tolerance towards undesired substances in molasses, U. maydis is better equipped for the process on the alternative feedstock molasses than U. cynodontis. The established process with U. maydis reached competitive yields of up to 0.38 g g−1 and a titre of more than 37 g L−1. This shows that an efficient and cost-effective itaconate production process is generally feasible using U. maydis, which has the potential to greatly increase the sustainability of industrial itaconate production. • Itaconate production from molasses with engineered Ustilago sp. • High invertase activity for sucrose hydrolysis is key to process efficiency • U. maydis shows superior sucrose conversion and raw material tolerance compared to U. cynodontis. • Fed-batch process with molasses achieves competitive itaconate yields and titres [ABSTRACT FROM AUTHOR]

The growing global market for industrial enzymes has led to a constant search for efficient, cost-effective methods for their production. This study reports the production of invertase using inexpensive and readily available agro-materials. Starch-digesting enzymes extracted from malted unkilned sorghum were used to hydrolyze cassava starch supplemented with 2% whole soybean. The production of intracellular invertase by Saccharomyces cerevisiae OL629078.1 in cassava-soybean and yeast sucrose broth was compared. The purification and characterization of invertase produced using the low-cost medium were also reported. The results showed that there was a 4.1-fold increase in the units of invertase produced in cassava-soybean medium (318.605 U/mg) compared to yeast sucrose broth medium (77.6 U/mg). The invertase produced was purified by chromatographic methods up to 5.53-fold with a recovery of 62.6%. Estimation of the molecular weight with gel filtration indicated a molecular weight of 118 kDa. The enzyme demonstrated its maximum activity at 50 °C and there was no decrease in its activity following a 1-h incubation at this temperature. At a pH of 5.0, the enzyme demonstrated optimal activity and it maintained over 60% of its activity in the acid range (pH 3–6). The Michalis-Menten constants Km and Vmax of intracellular invertase were 5.85 ± 1.715 mM and 6.472 ± 2.099 U/mg, respectively. These results suggest that Saccharomyces cerevisiae grown on cassava-soybean is a viable, cost-effective alternative for commercial invertase production, which can be explored for biotechnological processes. [ABSTRACT FROM AUTHOR]

Black rot disease, caused by Ceratocystis fimbriata Ellis & Halsted, severely affects both plant growth and post-harvest storage of sweet potatoes. Invertase (INV) enzymes play essential roles in hydrolyzing sucrose into glucose and fructose and participate in the regulation of plant defense responses. However, little is known about the functions of INV in the growth and responses to black rot disease in sweet potato. In this study, we identified and characterized an INV-like gene, named IbINV, from sweet potato. IbINV contained a pectin methylesterase-conserved domain. IbINV transcripts were most abundant in the stem and were significantly induced in response to C. fimbriata, salicylic acid, and jasmonic acid treatments. Overexpressing IbINV in sweet potato (OEV plants) led to vigorous growth and high resistance to black rot disease, while the down-regulation of IbINV by RNA interference (RiV plants) resulted in reduced plant growth and high sensitivity to black rot disease. Furthermore, OEV plants contained a decreased sucrose content and increased hexoses content, which might be responsible for the increased INV activities; not surprisingly, RiV plants showed the opposite effects. Taken together, these results indicate that IbINV positively regulates plant growth and black rot disease resistance in sweet potato, mainly by modulating sugar metabolism. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Bioprocess Engineering is the property of Chinese Journal of Bioprocess Engineering Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agronomica Sinica is the property of Crop Science Society of China and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Acta Agronomica Sinica is the property of Crop Science Society of China and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract The food enzyme β‐fructofuranosidase (β‐d‐fructofuranoside fructohydrolase; EC 3.2.1.26) is produced with the non‐genetically modified Saccharomyces cerevisiae strain NCYC R693 by Kerry Ingredients & Flavours Ltd. The production strain meets the requirements for the qualified presumption of safety (QPS) approach. The food enzyme is intended to be used in four food manufacturing processes. The dietary exposure to the food enzyme–total organic solids (TOS) was estimated to be up to 2.485 mg TOS/kg body weight per day in European populations. As the production strain qualifies for the QPS approach of safety assessment and no issue of concern arising from the production process of the food enzyme were identified, the Panel considered that no toxicological studies other than the assessment of allergenicity were necessary. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and one match with a tomato allergen was found. The Panel considered that the risk of allergic reactions upon dietary exposure to this food enzyme, particularly in individuals sensitised to tomato, cannot be excluded. However, the likelihood of allergic reactions is expected not to exceed the likelihood of allergic reactions to tomato. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Late spring cold (LSC) occurred in the reproductive period of wheat impairs spike and floret differentiation during the reproductive period, when young spikelets are very cold-sensitive. However, under LSC, the responses of wheat spikelets at various positions, leaves, and stems and the interactions between them at physiological levels remain unclear. In the present study, two-year treatments at terminal spikelet stage under two temperatures (2 °C, −2 °C) and durations (1, 2, and 3 days) were imposed in an artificial climate chamber to compare the effects of LSC on grain number and yield in the wheat cultivars Yannong 19 (YN19, cold-tolerant) and Xinmai 26 (XM26, cold-sensitive). The night temperature regimes were designed to reproduce natural temperature variation. LSC delayed plant growth and inhibited spike and floret differentiation, leading to high yield losses in both cultivars. LSC reduced dry matter accumulation (DMA, g) in spikes, stems, and leaves, reducing the DMA ratios of the spike to leaf and spike to stem. Plant cell wall invertase (CWINV) activity increased in upper and basal spikelets in YN19, whereas CWINV increased in middle spikelets in XM26. Under LSC, soluble sugar and glucose were transported and distributed mainly in upper and basal spikelets for glume and rachis development, so that spike development was relatively complete in YN19, whereas the upper and basal spikelets were severely damaged and most of the glumes in middle spikelets were relatively completely developed in XM26, resulting in pollen abortion mainly in upper and basal spikelets. The development of glumes and rachides was influenced and grain number per spike was decreased after LSC, with kernels present mainly in middle spikelets. Overall, reduced total DMA and dry matter partitioning to spikes under LSC results in poor spikelet development, leading to high losses of grain yield.

Sugar beet molasses is a low-value byproduct from the sugar industry. It contains significant amounts of sucrose (approx. 50% (w/w)), which can be used for many different applications, for example, as feedstock for the production of fuel (as ethanol) and biobased chemicals such as 5-hydoxymethyl furfural (HMF). To produce platform chemicals, sucrose is hydrolyzed into its monomeric C6 sugars: glucose and fructose. When comparing the hydrolysis rates of molasses with a pure sucrose solution, the specific reaction rate is much slower (Qp/x,60min = 93 and 70 gprod L−1 h−1 gcell−1 for pure sucrose and crude molasses, respectively) at the same sucrose concentration (300 g/L) and process conditions. To clarify why molasses inhibits the enzymatic hydrolysis rate, the influence of its viscosity and inorganic and organic composition was investigated. Also, the effects of molasses and treated molasses on pure enzymes, invertase (from Saccharomyces cerevisiae, 0.05 mg/mL), compared with hydrolysis using whole cells of Baker’s yeast (3 mg/mL), were tested. The results indicate an inhibitory effect of potassium (Qp/x,60min = 76 gprod L−1 h−1 gcell−1), generally at high salt concentrations (Qp/x,60min = 67 gprod L−1 h−1 gcell−1), which could be correlated to the solution’s high salt concentrations and possibly the synergistic effects of different ions when applying concentrations that were four times that in the molasses. Also, the viscosity and sucrose purity seem to have an effect, where pure sucrose solutions and thick juice from the sugar mill yielded higher hydrolysis rates (Qp/x,60min = 97 gprod L−1 h−1 gcell−1) than molasses-type solutions with a higher viscosity (Qp/x,60min = 70–74 gprod L−1 h−1 gcell−1). Attempting to further understand the effects of different components on the invertase activity, an in silico investigation was performed, indicating that high salt concentrations affected the binding of sucrose to the active site of the enzyme, which can result in a lower reaction rate. This knowledge is important for future scale-up of the hydrolysis process, since reduced hydrolysis rates require larger volumes to provide a certain productivity, requiring larger process equipment and thereby higher investment costs.

Studying the thermal stress effect on sucrose-metabolizing enzymes in sugarcane is of great importance for understanding acclimation to thermal stress. In this study, two varieties, S2003-US-633 and SPF-238, were grown at three different temperatures ( ± 2°C): 30°C as a control, 45°C for various episodes of high temperature treatments and recovery conditions at 24, 48 and 72 hours. Data showed that reducing sugar content increased until the grand growth stage but sharply declined at the maturity stage in both cultivars. On the other hand, sucrose is enhanced only at the maturity stage. The expression of all invertase isozymes declined prominently; however, the expression of SPS was high at the maturity stage. Hence, the sucrose accumulation in mature cane was due to increased SPS activity while decreased invertase isozymes (vacuolar, cytoplasmic and cell wall) activities at maturity stage in both cultivars. Heat shock decreased the sucrose metabolizing enzymes, sucrose content and sugar recovery rate in both cultivars. In contrast, heat-shock treatments induced maximum proline, MDA, H2O2 and EC in both cultivars. Notably, this is the first report of diverse invertase isozyme molecular weight proteins, such as those with 67, 134 and 160 kDa, produced under heat stress, suggesting that these enzymes have varied activities at different developmental stages. Overall, S2003-US-633 performs better than the cultivar SPF-238 under heat stress conditions at all development stages, with increased sucrose content, enzyme expression, proline and sugar recovery rate. This work will provide a new avenue regarding sugarcane molecular breeding programs with respect to thermal stress. [ABSTRACT FROM AUTHOR]

Inulinases are enzymes of great interest in the food industry, especially due to their application in the synthesis of fructose and fructo-oligosaccharides. Moreover, some inulinases (I) also present invertase activity (S), making them useful for sucrose hydrolysis processes. In the present study, the production of inulinase by Aspergillus niger URM5741 was evaluated and optimized using two statistical approaches. First, the composition of the cultivation medium was determined through a simplex centroid mixture design, followed by the selection of optimal fermentation conditions using the Box–Behnken design. Based on these experimental designs, the maximum activities of inulinase (16.68 U mL−1) and invertase (27.80 U mL−1) were achieved using a mixture of wheat, soy, and oat brans (5 g), along with 2.5% inulin and 40% moisture. The inulinase exhibited optimum temperature and pH of 60 °C and 4.0, respectively, displayed a high affinity for both substrates, as evidenced by very-low Michaelis constant values (1.07–1.54 mM). A relative thermostability was observed at 55–60 °C as indicated by half-lives values (I: 169.06–137.27 min; S: 173.29–141.52 min) and D-values (I: 561.61–456.00 min; S: 575.65–470.11 min) which were further confirmed by the high activation energy (123.01 and 143.29 kJ mol−1). The enzyme demonstrated favorable results in terms of inulin and sucrose hydrolysis, being a maximum release of reducing sugars of 6.04 and 15.80 g L−1, respectively. These results indicate that the sequential statistical approach proved to be beneficial to produce inulinase by A. niger URM5741, with the obtained enzyme considered promising for long-term industrial applications. [ABSTRACT FROM AUTHOR]

Invertase, an industrially significant glycoenzyme, was purified from baker's yeast using poly (2-Hydroxyethyl methacrylate) [PHema-Pba] cryogels functionalized with boronic acid. At subzero temperatures, PHema-Pba cryogels were synthesized and characterized using swelling tests, scanning electron microscopy, and Fourier-transform infrared spectroscopy. The surface area of the PHema-Pba cryogels was 14 m2/g with a swelling ratio of 88.3% and macroporosity of 72%. The interconnected macropores of PHema-Pba cryogels were shown via scanning electron microscopy. Invertase binding capacity of PHema-Pba cryogel was evaluated by binding studies in different pH, temperature, and interaction time conditions and the maximum Invertase binding of PHema-Pba cryogel was found as 15.2 mg/g. and 23.7 fold Invertase purification was achieved from baker's yeast using PHema-Pba cryogels. The results show that PHema-Pba cryogels have high Invertase binding capacity and may be used as an alternative method for enzyme purification via boronate affinity systems. [ABSTRACT FROM AUTHOR]

Bud dormancy enables deciduous fruit trees to endure unfavorable conditions during winter, and considerably impacts growth and reproduction in spring. We investigated acid invertase activities during dormancy release until bud break under natural (orchard) conditions in two consecutive years. Our aim was to relate the activity patterns to the developmental progression and to air temperature, which is a major factor influencing the developmental processes at that stage. The enzyme assays were performed on extracts from leaf buds of the cultivar Idared, sampled from early March to April in the years 2020 and 2021. The air temperature was continually monitored during the observation periods. cwINV activity showed a trend of slight increase at the earlier developmental stages and sharply increased during bud break in both years. cwINV is known to contribute to organ sink strength. Its up-regulation may, therefore, be related to the increasing developmental demand for carbohydrates in apple leaf buds during dormancy release until bud break. vacINV activity was relatively constant at the earlier stages and also showed a pronounced increase in activity during bud break in both years. However, in both years, we observed drops in vacINV activity following cold spells. vacINV activity has been associated with growth via cell elongation through the regulation of cell turgor and may, thus, be involved in bud break. Therefore, we suggest that the down-regulation of vacINV activity as a consequence of cold spells might contribute to a delay in bud break to protect young leaf tissues from exposure to cold stress conditions. [ABSTRACT FROM AUTHOR]