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Abstract Background The encapsulation of metagenome-derived multi-enzymes presents a novel approach to improving poultry feed by enhancing nutrient availability and reducing anti-nutritional factors. By integrating and encapsulated enzymes such as carbohydrate-hydrolyzing enzymes, protease, lipase, and laccase into feed formulations, this method not only improves feed digestibility but also potentially contributes to animal health and productivity through antimicrobial properties. Results This study investigates the encapsulation of metagenome-derived enzymes, including carbohydrate-hydrolyzing enzymes, protease, lipase, and laccase, using Arabic and Guar gums as encapsulating agents. The encapsulated multi-enzymes exhibited significant antimicrobial activity, achieving a 92.54% inhibition rate against Escherichia coli at a concentration of 6 U/mL. Fluorescence tracking with FITC-labeled enzymes confirmed efficient encapsulation and distribution, while physical characterization, including moisture content and solubility assessments, along with Atomic Force Microscopy (AFM) imaging, validated successful encapsulation. The encapsulated enzymes also effectively hydrolyzed poultry feed, leading to an increase in phenolic content and antioxidant activity, as confirmed by 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Conclusions The encapsulated multi-enzymes improved the overall feed quality by increasing reducing sugars and enhancing physical properties such as solubility and water-holding capacity. The encapsulated multi-enzymes improved the overall feed quality by increasing reducing sugars, antioxidant activity and enhancing physical properties such as solubility and water-holding capacity. Scanning Electron Microscopy (SEM) and Fourier-Transform Infrared Spectroscopy (FTIR) analyses confirmed the enzymatic breakdown of the feed structure. These results suggest that supplementing poultry feed with encapsulated multi-enzymes can enhance its physical, nutritional, and functional properties, leading to improved digestibility and overall feed quality.

Background: The encapsulation of metagenome-derived multi-enzymes presents a novel approach to improving poultry feed by enhancing nutrient availability and reducing anti-nutritional factors. By integrating and encapsulated enzymes such as carbohydrate-hydrolyzing enzymes, protease, lipase, and laccase into feed formulations, this method not only improves feed digestibility but also potentially contributes to animal health and productivity through antimicrobial properties. Results: This study investigates the encapsulation of metagenome-derived enzymes, including carbohydrate-hydrolyzing enzymes, protease, lipase, and laccase, using Arabic and Guar gums as encapsulating agents. The encapsulated multi-enzymes exhibited significant antimicrobial activity, achieving a 92.54% inhibition rate against Escherichia coli at a concentration of 6 U/mL. Fluorescence tracking with FITC-labeled enzymes confirmed efficient encapsulation and distribution, while physical characterization, including moisture content and solubility assessments, along with Atomic Force Microscopy (AFM) imaging, validated successful encapsulation. The encapsulated enzymes also effectively hydrolyzed poultry feed, leading to an increase in phenolic content and antioxidant activity, as confirmed by 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Conclusions: The encapsulated multi-enzymes improved the overall feed quality by increasing reducing sugars and enhancing physical properties such as solubility and water-holding capacity. The encapsulated multi-enzymes improved the overall feed quality by increasing reducing sugars, antioxidant activity and enhancing physical properties such as solubility and water-holding capacity. Scanning Electron Microscopy (SEM) and Fourier-Transform Infrared Spectroscopy (FTIR) analyses confirmed the enzymatic breakdown of the feed structure. These results suggest that supplementing poultry feed with encapsulated multi-enzymes can enhance its physical, nutritional, and functional properties, leading to improved digestibility and overall feed quality. [ABSTRACT FROM AUTHOR]

Background: Enzyme combinations, particularly phytase (PHY) with various carbohydrases and proteases, are utilized in commercial broiler production to enhance nutrient and energy bioavailability. Objective: A feeding study was undertaken to determine whether the efficiency of an Escherichia coli–derived PHY and a feed enzyme complex (FEC) derived from Bacillus spp. containing carbohydrase and protease as main activities in broiler chickens is dependent on diet quality. A total of 900 male one‐day‐old broiler chickens (Ross 308) were assigned to a 2 × 3 factorial arrangement of the treatments with 2 different nutrient density diets, standard nutrient diet (SN diet) and a low‐nutrient diet (LN diet; −100 kcal/kg for AMEn and −5% for crude protein [CP] and limiting amino acids), and 3 enzyme treatments (control [no enzymes], PHY and PHY + FEC). Each treatment group was composed of 6 replicates of 25 birds each. Results: The LN diet caused a decrease in performance index, tibia length and diameter, tibia calcium content and jejunal villus surface area (VSA). The interaction effects between diet and enzyme supplementation were observed (p < 0.05) on overall average daily gain (ADG), performance index, tibia ash content and jejunal villus height (VH) and VSA, with the favourable benefits of PHY + FEC treatment being more pronounced in the LN diets. Regardless of dietary nutrient density, supplementation with PHY alone or combined with FEC enhanced (p < 0.05) final body weight, overall ADG and jejunal villus height (VH)/crypt depth, with the highest values observed in the PHY + FEC group. The PHY + FEC treatment also improved (p < 0.05) overall feed conversion ratio, apparent ileal digestibility of dry matter, organic matter, CP, and energy, and tibia phosphorus content compared to the control treatment. Conclusions: The results indicate that the simultaneous addition of PHY and FEC to the LN diets improved the growth rate, bone mineralization and gut morphology. [ABSTRACT FROM AUTHOR]

A study determined the effects of supplementing corn-based diets for weaned pigs with multi-enzymes on growth performance, apparent total tract digestibility (ATTD) of nutrients, fecal score, and fecal microbial composition. A total of 132 pigs (initial body weight = 7.23 kg) that had been weaned at 21 d of age and fed a drug-free nursery diet for 7 d were housed in 33 pens of 4 barrows or gilts, blocked by body weight and gender, and fed 3 experimental diets at 11 pens per diet. The diets were corn-based diet without or with multi-enzyme A or B. Multi-enzyme A supplied 4,000 U of xylanase, 150 U of β-glucanase, 3,500 U of protease, and 1,500 U of amylase per kilogram of diet. Multi-enzyme B was the same as multi-enzyme A except that it supplied amylase at 150 U/kg, and that its source of amylase was different from that of multi-enzyme A. All diets contained phytase at 1,000 U/kg. The diets were fed for 35 d in 2 phases; phase 1 for the first 14 d and phase 2 for the last 21 d of the trial. Fecal score was determined daily during the first 7 d of the trial. Fecal samples were collected from rectum of 1 pig per pen on days 2, 7, 14, and 35 of the trial for determining bacterial composition. Also, fresh fecal samples were collected from each pen on days 41 and 42 to determine ATTD of nutrients. Multi-enzyme B increased (P < 0.05) average daily gain (ADG) for phases 1 and 2. For the overall study period, multi-enzyme B increased (P < 0.05) ADG from 262 to 313 g, and average daily feed intake (ADFI) from 419 to 504 g. Multi-enzyme A increased (P < 0.05) overall ADG from 262 to 290 g, but did not affect ADFI. Multi-enzyme A or B did not affect ATTD of gross energy, but increased (P < 0.05) the ATTD of ether extract from 30% to 36% or 37%, respectively. Multi-enzyme A did not affect fecal score; however, multi-enzyme B tended to decrease (P = 0.09) fecal score, implying that it tended to decrease diarrhea. Firmicutes were the most abundant phylum of fecal bacteria (its relative abundance ranged from 58% to 72%). Bacteroidetes and Actinobacteria were the 2nd and 3rd most abundant phyla of fecal bacteria. Neither multi-enzyme affected fecal bacterial composition. In conclusion, the addition of multi-enzyme A or B to phytase-supplemented corn-based diet for weaned pigs can improve their growth performance and fat digestibility. However, multi-enzyme B was more effective than multi-enzyme A in terms of improving the growth performance of weaned pigs fed corn-based diet.

Objective The present study examined the effects of exogenous emulsifiers and multi-enzyme supplementation into a low energy density diet on growth performance, visceral organ parameters, blood metabolites, ileal morphology, and nutrient digestibility in broiler chickens from hatch to 21 days. Methods One hundred and sixty-eight one-day-old Ross 308 broiler chickens were allocated in a completely randomized design to 24 pens and each pen was assigned to one of four dietary treatments to give six replications with seven chickens in a cage. Dietary treatments were: i) positive control with standard energy level (PC); ii) negative control with 100 kcal/kg lower energy of the standard level (NC); iii) NC diet supplemented 0.05% calcium stearoyl-2 lactylate as an emulsifier (NC+E); and iv) NC diet supplemented with both 0.05% calcium stearoyl-2 lactylate and 0.05% multi-enzyme (NC+E+M). Corn and soybean meal-based control diets containing vegetable oil were formulated to meet the Ross 308 nutrition specification. Chickens were fed ad-libitum with the treatment diets and sampling was conducted on day 21. Results Our results revealed that emulsifier and multi-enzyme supplementation into NC diets improved (p0.05) visceral organ weight, blood metabolites, and intestinal morphology in broiler chickens fed NC diets. Conclusion Supplementation of emulsifier and multi-enzyme in the NC diet would support improving growth performance in young broiler chickens with improved feed efficiency and increased nutrient digestibility thereby curtailing the negative impact of energy reduction in the diets.

Bacillus spp. have gained increasing recognition as an option to use as antimicrobial growth promoters, which are characterized by producing various enzymes and antimicrobial compounds. The present study was undertaken to screen and evaluate a Bacillus strain with the multi-enzyme production property for poultry production. LB-Y-1, screened from the intestines of healthy animals, was revealed to be a Bacillus velezensis by the morphological, biochemical, and molecular characterization. The strain was screened out by a specific screening program, possessed excellent multienzyme production potential, including protease, cellulase, and phytase. Moreover, the strain also exhibited amylolytic and lipolytic activity in vitro. The dietary LB-Y-1 supplementation improved growth performance and tibia mineralization in chicken broilers, and increased serum albumin and serum total protein at 21 days of age (p < 0.05). Besides, LB-Y-1 enhanced the activity of serum alkaline phosphatase and digestive enzyme in broilers at 21 and 42 days of age (p < 0.05). Analysis of intestinal microbiota showed that a higher community richness (Chao1 index) and diversity (Shannon index) in the LB-Y-1 supplemented compared with the CON group. PCoA analysis showed that the community composition and structure were distinctly different between the CON and LB-Y-1 group. The beneficial genera such as Parasutterella and Rikenellaceae were abundant, while the opportunistic pathogen such as Escherichia-Shigella were reduced in the LB-Y-1 supplemented group (p < 0.05). Collectively, LB-Y-1 can be considered as a potential strain for further utilization in direct-fed microbial or starter culture for fermentation. [ABSTRACT FROM AUTHOR]

Objective: The present study examined the effects of exogenous emulsifiers and multienzyme supplementation into a low energy density diet on growth performance, visceral organ parameters, blood metabolites, ileal morphology, and nutrient digestibility in broiler chickens from hatch to 21 days. Methods: One hundred and sixty-eight one-day-old Ross 308 broiler chickens were allocated in a completely randomized design to 24 pens and each pen was assigned to one of four dietary treatments to give six replications with seven chickens in a cage. Dietary treatments were: i) positive control with standard energy level (PC); ii) negative control with 100 kcal/kg lower energy of the standard level (NC); iii) NC diet supplemented 0.05% calcium stearoyl-2 lactylate as an emulsifier (NC+E); and iv) NC diet supplemented with both 0.05% calcium stearoyl-2 lactylate and 0.05% multi-enzyme (NC+E+M). Corn and soybean meal-based control diets containing vegetable oil were formulated to meet the Ross 308 nutrition specification. Chickens were fed ad-libitum with the treatment diets and sampling was conducted on day 21. Results: Our results revealed that emulsifier and multi-enzyme supplementation into NC diets improved (p<0.05) feed efficiency of the broiler chickens compared to the broiler chickens fed NC diets from hatch to 21 days. Supplementation of emulsifier and multienzyme into NC diet improved (p<0.05) nutrient digestibility of the broiler chickens. However, emulsifier and multi-enzymesupplementation into diet did not influence (p>0.05) visceral organ weight, blood metabolites, and intestinal morphology in broiler chickens fed NC diets. Conclusion: Supplementation of emulsifier and multi-enzyme in the NC diet would support improving growth performance in young broiler chickens with improved feed efficiency and increased nutrient digestibility thereby curtailing the negative impact of energy reduction in the diets. [ABSTRACT FROM AUTHOR]

This research aimed to study the impact of supplementation of three multi-enzyme levels (0, 0.1, and 0.2% of feed) and two levels of dietary treatments [standard diet (SD) and low-density diet (LDD)] on growth performance, carcass traits, digestibility, and meat quality of broilers from 1 to 38 days of age. A total of 216 1-day-old Arbor Acres broiler chicks were randomly assigned to a factorial experiment (2 × 3) comprising six dietary treatments, each with six replicates and each replicate with six chickens. The results showed that the LDD significantly reduced body weight gain by 5.0%, compared with the SD. Multi-enzymes significantly improved body weight gain and the production index (PI) relative to the SD. The feed conversion ratio was significantly enhanced with increased multi-enzymes from 1 to 21 days. A significant relation between the multi-enzyme concentration and type of dietary treatment was observed in body weight gain and feed conversion ratio from 1 to 21 days of age. Nitrogen-free extract digestibility was significantly increased by using the SD diet compared with using the LDD. Multi-enzyme supplementation improved the digestibility of dry matter, crude protein, crude fiber, and nitrogen-free extract in the LDD. A significant relationship was found between the multi-enzyme concentration and type of dietary treatment on the pancreas, liver, and intestinal length percentages. The meat dry matter concentration was significantly higher in the LDD group than in the SD group. The low-density diet significantly reduced the total revenue compared with the SD, whereas broilers fed the SD recorded significantly higher total revenue and economic efficiency than those fed the LDD. The low-density diet significantly increased economic efficiency compared with the SD. Multi-enzymes significantly increased the total revenue, net revenue, and economic efficiency than the standard set. In conclusion, using multi-enzymes in broiler diets improved body weight gain. The LDD with multi-enzymes showed enhanced body weight gain compared with the SD without multi-enzymes.

We propose a co-immobilized chemo-enzyme cascade system to mitigate random intermediate diffusion from the mixture of individual immobilized catalysts and achieve a one-pot reaction of multi-enzyme and reductant. Catalyzed by lipase and lipoxygenase, unsaturated lipid hydroperoxides (HPOs) were synthesized. 13(S)-hydroperoxy-9Z, 11E-octadecadienoic acid (13-HPODE), one compound of HPOs, was subsequently reduced to 13(S)-hydroxy-9Z, 11E-octadecadienoic acid (13-HODE) by cysteine. Upon the optimized conditions, 75.28 mg of 13-HPODE and 4.01 mg of 13-HODE were produced from per milliliter of oil. The co-immobilized catalysts exhibited improved yield compared to the mixture of individually immobilized catalysts. Moreover, it demonstrated satisfactory durability and recyclability, maintaining a relative HPOs yield of 78.5% after 5 cycles. This work has achieved the co-immobilization of lipase, lipoxygenase and the reductant cysteine for the first time, successfully applying it to the conversion of soybean oil into 13-HODE. It offers a technological platform for transforming various oils into high-value products. [Display omitted] • The formation of metal-amino acid frameworks was facilitated by the addition of PVP and L-cysteine. • In a one-pot cascade reaction, soybean oil was successfully converted into 13-HPODE and 13-HODE. • The in-situ co-immobilization of lipase, lipoxygenase and the reductant L-cysteine has been accomplished within MOF. [ABSTRACT FROM AUTHOR]

[Display omitted] • Biomass-derived bioethanol is used to produce acetoin. • An efficient magnetic multienzyme cascade system is developed. • Immobilized NAD+ enhances the cofactor regeneration. • Maximum conversion rate of 90.4% is obtained under cofactor regeneration. • Magnetic co-immobilization system retains better operational stability. Upgrading biomass-derived bioethanol to higher-order alcohols using conventional biotechnological approaches is challenging. Herein, a novel, magnetic metal–organic-framework-based cofactor regeneration system was developed using ethanol dehydrogenase (EtDH:D46G), NADH oxidase (NOX), formolase (FLS:L482S), and nicotinamide adenine dinucleotide (NAD+) for converting rice straw-derived bioethanol to acetoin. A magnetic zeolitic imidazolate framework-8@Fe 3 O 4 /NAD+ (ZIF-8@Fe 3 O 4 /NAD+) regeneration system for cell-free cascade reactions was introduced and used to encapsulate EtDH:D46G, NOX, and FLS:L482S (ENF). ZIF-8@Fe 3 O 4 /NAD+ENF created an efficient microenvironment for three-step enzyme cascades. Under the optimized conditions, the yield of acetoin from 100 mM bioethanol using ZIF-8@Fe 3 O 4 /NAD+ENF was 90.4 %. The regeneration system showed 97.1 % thermostability at 50 °C. The free enzymes retained only 16.3 % residual conversion, compared with 91.2 % for ZIF-8@Fe 3 O 4 /NAD+ENF after ten cycles. The magnetic metal–organic-framework-based cofactor regeneration system is suitable for enzymatic cascade biotransformations and can be extended to other cascade systems for potential biotechnological applications. [ABSTRACT FROM AUTHOR]

Simple Summary: Insufficient energy and nutrient ingestion of primiparous lactating sows causes excess body weight loss and oxidative stress and compromises piglet growth. Two strategies to enhance the daily intake of dietary energy and nutrients were compared with a standard lactation diet to understand the potential modes of action. We found that feeding on either high-quality or flavor plus multi-enzyme diets both improved sow feed digestibility and consequently increased the growth of piglets. In addition, the flavor plus multi-enzyme diet also improved the antioxidant capacity and health of sows. The data suggest that dietary supplementation with flavor and multi-enzymes may be more promising than a high-quality diet from a health and economic perspective like enhancing utilization of cereal byproducts and thus reducing expenditure of corn and soybeans. This study was aimed to explore how a high-quality diet or a flavor plus multi-enzyme diet affects the feed intake, nutrient digestibility and antioxidation capacity of lactating sows and the growth of their progeny. Thirty primiparous sows were randomly assigned to three treatments from d 2 of lactation until weaning (d 21): control (CON), with a basal diet; high quality (HQ), with 200 kcal/kg higher net energy than CON; or the CON diet supplemented with 500 mg/kg flavor and 100 mg/kg multi-enzymes (F + E). Sows fed with the HQ or F + E diets improved piglets' live weight (p < 0.05) and average daily weight gain (p < 0.10), litter weight gain (p < 0.10) and piglet growth to milk yield ratio (p < 0.10). Compared with CON, the HQ and F + E groups increased the digestibility of ether extract, ash, neutral detergent fiber, crude fiber and phosphorus (p < 0.10), and the HQ group also increased dry matter, gross energy, crude protein, acid detergent fiber and energy intake (p < 0.05). Compared with CON, the F + E group decreased serum urea nitrogen and aspartate aminotransferase (p < 0.05) and enhanced superoxide dismutase, catalase and glutathione peroxidase, but it decreased malondialdehyde in milk supernatant (p < 0.05). [ABSTRACT FROM AUTHOR]

Polyelectrolyte-doped microcapsules (PDM) was fabricated by coaxial electrospray of a mixture of glycerol and water containing 10 mg/mL cationic polyelectrolyte poly(allylamine hydrochloride) (PAH) fed as the core phase solution, and a N,N-dimethylacetylamide solution of 10 wt% polyurethane fed as the shell phase solution. Multi-enzyme system involving Candida Antarctica lipase B (CALB), glucose oxidase (GOD), and horseradish peroxidase (HRP) for cascade reaction was assembled in the PDM at three different places, namely, surface, shell, and lumen. Placing of enzyme inside aqueous lumen of the PDM was realized by in situ encapsulation through adding the enzyme in the core-phase solution for coaxial electrospray. By ion-pairing of enzyme with cationic surfactant CTAB, an organic soluble enzyme-CTAB complex was prepared, so that in situ embedding of enzyme in the shell of the PDM was realized by adding it into the shell phase solution. Surface attachment of enzymes was achieved by layer-by-layer (LbL) technology, which is based on the ion-exchange interactions between oppositely charged enzymes and PAH that was doped in PDM. The enzyme-decorated microcapsule was then studied as a micro-bioreactor, in which 1-Oxododecyla-α-glucopyranoside was converted by CALB to glucose, which was oxidised by GOD to gluconolactone in a second step. The hydrogen peroxide produced was then used by HRP to oxidize ABTS to form coloured radical cation ABTS•+ for activity analysis. The successful fabrication of the PDM and precise localization of enzymes in the PDM by different strategies were fully characterized. By varying the immobilization strategy, totally six PDM bioreactors with three enzymes precisely positional assembled in different strategies were constructed and their activities for the cascade reaction were investigated and compared. The PDM micro-bioreactor prepared by novel electrospray technologies provide a smart platform for positional assembly of multi-enzyme cascade reaction in a precise and well-controlled manner.

Laying and reproductive performance, egg quality, and disease resistance of hens decrease during the late laying period. Exogenous enzymes promote nutrient digestibility and utilization and improve the intestinal environment. However, the specific regulation of the gut microbiome and metabolome by exogenous enzymes remains unelucidated. This study was conducted to evaluate effects of dietary multi-enzyme supplementation on egg and reproductive performance, egg quality, ileum microbiome, and metabolome of breeders. Here, 224 Hy-Line Brown breeding hens (55 weeks old) were randomly allocated to two groups: dietary controls fed basal diet (DC), and test hens fed 0.2 g/kg corn enzyme diet (CE). Serum levels of total protein, globulin, immunoglobulin Y, and antibodies against the Newcastle disease virus and avian influenza H9 strain were significantly increased (p < 0.05). Egg albumen height, Haugh unit, and fertilization and hatching rates were also significantly increased (p < 0.05) in the CE-fed group. 16S rRNA sequence analysis showed that CE strongly affected both α- and β-diversity of the ileal microbiota. LEfSe analysis revealed that the potentially beneficial genera Lactobacillus , Enterococcus , Faecalicoccus , and Streptococcus were enriched as biomarkers in the CE-fed group. Microbial functional analysis revealed that the functional genes associated with harmful-substance biodegradation was significantly increased in the CE-fed group. Additionally, Spearman correlation analysis indicated that changes in microbial genera were correlated with differential metabolites. In summary, dietary multi-enzyme addition can improve egg quality, humoral immunity, and reproductive performance and regulate the intestinal microbiome and metabolome in breeders. Therefore, multi-enzymes could be used as feed additive to extend breeder service life. [ABSTRACT FROM AUTHOR]

Laying and reproductive performance, egg quality, and disease resistance of hens decrease during the late laying period. Exogenous enzymes promote nutrient digestibility and utilization and improve the intestinal environment. However, the specific regulation of the gut microbiome and metabolome by exogenous enzymes remains unelucidated. This study was conducted to evaluate effects of dietary multi-enzyme supplementation on egg and reproductive performance, egg quality, ileum microbiome, and metabolome of breeders. Here, 224 Hy-Line Brown breeding hens (55 weeks old) were randomly allocated to two groups: dietary controls fed basal diet (DC), and test hens fed 0.2 g/kg corn enzyme diet (CE). Serum levels of total protein, globulin, immunoglobulin Y, and antibodies against the Newcastle disease virus and avian influenza H9 strain were significantly increased (p < 0.05). Egg albumen height, Haugh unit, and fertilization and hatching rates were also significantly increased (p < 0.05) in the CE-fed group. 16S rRNA sequence analysis showed that CE strongly affected both α- and β-diversity of the ileal microbiota. LEfSe analysis revealed that the potentially beneficial genera Lactobacillus, Enterococcus, Faecalicoccus, and Streptococcus were enriched as biomarkers in the CE-fed group. Microbial functional analysis revealed that the functional genes associated with harmful-substance biodegradation was significantly increased in the CE-fed group. Additionally, Spearman correlation analysis indicated that changes in microbial genera were correlated with differential metabolites. In summary, dietary multi-enzyme addition can improve egg quality, humoral immunity, and reproductive performance and regulate the intestinal microbiome and metabolome in breeders. Therefore, multi-enzymes could be used as feed additive to extend breeder service life.

A study was undertaken to determine the effects of supplemental multicarbohydrase (MC) and butyrate glycerides (BG) on productive performance, nutritional, and physiological responses in laying hens fed corn- or wheat-based diets during a 12-week production period (from 50–62 wk of age). The experiment consisted of a 2 × 2 × 2 factorial arrangement of the treatments with 2 different basal diets (corn- or wheat-based diets), 2 concentrations of MC (0 or 200 mg/kg of diet), and 2 concentrations of BG (0 or 2 g/kg of diet). Each treatment had 6 replicates with 8 hens each. The interactions among diet, MC, and BG were observed for egg production (P = 0.048), feed conversion ratio (P = 0.005), and ileal Escherichia coli count (P = 0.043), indicating that the effects of MC and BG on these responses were more marked when wheat-based diet was fed. A diet × MC interaction (P

Multi‐enzyme cascade reactions are indispensable in biotechnology and many industrial (bio)chemical processes. However, most natural enzymes have poor stability and reusability, and tend to inactivate in toxic media or high temperature, which significantly limit their broader applications. Metal‐organic frameworks (MOFs) are promising candidates for enzymes immobilization to produce nanocomposite structures that not only could shield the enzymes from harsh environments, but also facilitate selective diffusion of substrates and intermediates to the reactive site via their tailorable and ordered pore network. Multi‐enzyme cascade reactions in MOFs have recently attracted considerable attention. This Personal Account discusses the different strategies for multi‐enzyme‐MOF interfaces and their cutting‐edge applications from biosensing and catalytic nanomedicine to artificial/hybrid cells. At last, we provide a critical evaluation and future prospects to outline future research directions. [ABSTRACT FROM AUTHOR]

A study determined the effects of supplementing corn-based diets for weaned pigs with multi-enzymes on growth performance, apparent total tract digestibility (ATTD) of nutrients, fecal score, and fecal microbial composition. A total of 132 pigs (initial body weight = 7.23 kg) that had been weaned at 21 d of age and fed a drug-free nursery diet for 7 d were housed in 33 pens of 4 barrows or gilts, blocked by body weight and gender, and fed 3 experimental diets at 11 pens per diet. The diets were corn-based diet without or with multi-enzyme A or B. Multi-enzyme A supplied 4,000 U of xylanase, 150 U of β-glucanase, 3,500 U of protease, and 1,500 U of amylase per kilogram of diet. Multi-enzyme B was the same as multienzyme A except that it supplied amylase at 150 U/kg, and that its source of amylase was different from that of multienzyme A. All diets contained phytase at 1,000 U/kg. The diets were fed for 35 d in 2 phases; phase 1 for the first 14 d and phase 2 for the last 21 d of the trial. Fecal score was determined daily during the first 7 d of the trial. Fecal samples were collected from rectum of 1 pig per pen on days 2, 7, 14, and 35 of the trial for determining bacterial composition. Also, fresh fecal samples were collected from each pen on days 41 and 42 to determine ATTD of nutrients. Multi-enzyme B increased (P < 0.05) average daily gain (ADG) for phases 1 and 2. For the overall study period, multi-enzyme B increased (P < 0.05) ADG from 262 to 313 g, and average daily feed intake (ADFI) from 419 to 504 g. Multi-enzyme A increased (P < 0.05) overall ADG from 262 to 290 g, but did not affect ADFI. Multi-enzyme A or B did not affect ATTD of gross energy, but increased (P < 0.05) the ATTD of ether extract from 30% to 36% or 37%, respectively. Multi-enzyme A did not affect fecal score; however, multienzyme B tended to decrease (P = 0.09) fecal score, implying that it tended to decrease diarrhea. Firmicutes were the most abundant phylum of fecal bacteria (its relative abundance ranged from 58% to 72%). Bacteroidetes and Actinobacteria were the 2nd and 3rd most abundant phyla of fecal bacteria. Neither multi-enzyme affected fecal bacterial composition. In conclusion, the addition of multi-enzyme A or B to phytase-supplemented corn-based diet for weaned pigs can improve their growth performance and fat digestibility. However, multi-enzyme B was more effective than multi-enzyme A in terms of improving the growth performance of weaned pigs fed corn-based diet. [ABSTRACT FROM AUTHOR]

A total of 672 one-d-old Ross 308 (mixed gender) broiler chicks with an average initial body weight of 34.1 ± 0.2 g were used in a 28-d feeding trial to investigate the effect of supplementing low energy diets with an emulsifier or multi-enzyme complex on broiler. Chickens were allocated to one of six treatments with seven pens/treatment and 16 birds/pen. Treatments were (1) PC (basal diet), (2) NC (PC-100 kcal ME/kg), (3) L05 (NC + 0.05% emulsifier), (4) L10 (NC + 0.10% emulsifier), (5) E05 (NC + 0.05% multi-enzyme), and (6) E10 (NC + 0.10% multi-enzyme). Results showed that supplementing low energy diet with 0.10% multi-enzyme or 0.10% emulsifier improved (P

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Tyrosinase is a copper-containing metalloprotein that catalyzes the oxidation of tyrosine, in particular, L-DOPA to L-Dopaquinone, which are precursors of brown pigments in some wounded eukaryotic tissues. The present study focused on screening, production and characterization of tyrosinase from multi-enzyme producing Fusarium solani B1 and Fumago sp. A total of 25 strains were isolated from rotting wood samples and screened for hydrolytic and oxidative multi-enzyme potentials using different polymeric substrates. The two most consistent strains: Fusarium solani B1 and Fumago sp. B13 were further evaluated for tyrosinase production. Some media cultural parameters and physiological conditions were optimized in order to maximize tyrosinase production. Incubation of Fumago sp. B13 and Fusarium solani B1 for 96 and 144 h in medium containing 2 % and 0.2 % ratios of Glucose and NaNO3 with pH 6 and 7, respectively, was most suitable for tyrosinase production. Characterization of the partially purified tyrosinase from Fumago sp. B13 and Fusarium solani B1 exhibited optimal activities at pH 6-7, 30 °C, and 1 mM Cu2+, respectively, thereby suggesting their potentials for novel biotechnological applications. [ABSTRACT FROM AUTHOR]

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A study was conducted to determine effects of supplementing multi-enzyme on apparent ileal digestibility (AID) of energy and AA; and apparent total tract digestibility (ATTD) of energy for pigs fed low-energy and AA diets. Eight ileal-cannulated barrows (initial BW: 38.7 ± 2.75 kg) were fed four diets in a replicated 4 × 4 Latin square design to give 8 replicates per diet. Diets were positive control (PC) diet, negative control (NC) diet without or with multi-enzyme at 0.5 or 1.0 g/kg. The PC diet was formulated to meet or exceed NRC (2012) nutrient recommendations for grower pigs (25 to 50 kg), except for Ca and digestible P, which were lower than NRC (2012) recommendations by 0.13 and 0.17 percentage points, respectively, due to phytase supplementation at 1,000 FTU/kg. The NC diet was formulated to be lower in NE by 75 kcal/kg and standardized ileal digestible AA content by a mean of 3%. These reductions were achieved by partial replacement of corn and soybean meal (SBM) and complete replacement of soybean oil and monocalcium phosphate in PC diet with 25% corn distillers dried grains with solubles (DDGS) and 3.6% soybean hulls. Multi-enzyme at 1.0 g/kg supplied 1,900 U of xylanase, 300 U of β-glucanase, 1,300 U of cellulase, 11,500 U of amylase, 120 U of mannanase, 850 U of pectinase, 6,000 U of protease, and 700 U of invertase per kilogram of diet. The AID of GE, N, most AA, most component sugars of nonstarch polysaccharides (NSP) and P; ATTD of GE for PC diet was greater (P < 0.05) than those for NC diets. An increase in dietary level of multi-enzyme from 0 to 1.0 g/kg resulted in a linear increase (P < 0.05) in AID of Ile by 4.3%, and tended to linearly increase (P < 0.10) AID of Leu, Met, Phe, and Val by a mean of 3.4%. Increasing dietary multi-enzyme from 0 to 1.0 g/kg linearly increased (P < 0.05) AID of total NSP and P by 53.7% and 19.2%, respectively; ATTD of GE by 8.4% and DE and NE values by 8.8% and 8.2%, respectively; tended to linearly increase (P < 0.10) AID of GE by 8.1%. The NE values for NC diet with multi-enzyme at 1.0 g/kg tended to be greater (P < 0.10) than that for PC diet (2,337 vs. 2,222 kcal/kg of DM). In conclusion, multi-enzyme supplementation improved energy and nutrient digestibilities of a corn–SBM–corn DDGS-based diet, implying that the multi-enzyme fed in the current study can be used to enhance energy and nutrient utilization of low-energy AA diets for grower pigs. [ABSTRACT FROM AUTHOR]

A total of 672 one-d-old Ross 308 (mixed gender) broiler chicks with an average initial body weight of 34.1 ± 0.2 g were used in a 28-d feeding trial to investigate the effect of supplementing low energy diets with an emulsifier or multi-enzyme complex on broiler. Chickens were allocated to one of six treatments with seven pens/treatment and 16 birds/pen. Treatments were (1) PC (basal diet), (2) NC (PC-100 kcal ME/kg), (3) L05 (NC + 0.05% emulsifier), (4) L10 (NC + 0.10% emulsifier), (5) E05 (NC + 0.05% multi-enzyme), and (6) E10 (NC + 0.10% multi-enzyme). Results showed that supplementing low energy diet with 0.10% multi-enzyme or 0.10% emulsifier improved (P < .05) overall body weight gain. The relative weight of breast meat decreased (P < .05) by adding 0.10% multi-enzyme compared to chickens fed E05 diet. Feeding the chickens with E10 diet increased relative weight of abdominal fat. Chickens fed NC or L10 diets had the lowest (P < .05) relative weight of liver. The addition of emulsifier or multi-enzyme to the diets decreased drip loss (P < .05). Results indicated that supplementing low energy diet with multi-enzyme or emulsifier improved growth performance and alleviated negative effects of lowering dietary ME level on meat quality of broilers. [ABSTRACT FROM AUTHOR]

Nature is a source of inspiration for researchers and following the example of cells, the immobilization of several enzymes to produce chemicals of interest is a successful strategy. Co-immobilization is not a trivial issue and the choice of the most appropriate methodology and support is a challenge. Supports play a fundamental role in the immobilization process, several aspects regarding its chemical structure, its physical properties and other aspects as biodegradability and biocompatibility need to be carefully considered. Among natural polymers, chitin and chitosan have been tested as supports for enzyme and cells immobilization due to their unique properties. In this review, the production of d-amino acids using hydantoin hydrolysing biocatalyst (whole cells, crude cell extracts and isolated proteins) immobilized using chitin and chitosan is reported and compared with other supports. [ABSTRACT FROM AUTHOR]

This study evaluated the effects of multi-enzyme (xylanase, mannanase, β-glucanase, cellulase, pectinase, and amylase) supplementation on growth performance, nutrient digestibility, blood characteristics, relative organ weight, and breast muscle quality in broilers fed maize-wheat-soybean meal-based diets. A total of 800, one-day-old Ross 308 broiler chicks were randomly assigned to one of 4 treatments according body weight (body weight, 45.3 ± 0.72 g) in a 35-day trail. Each treatment had 10 replicate cages with 20 birds per replicate cage. Dietary treatment groups were: (1) CON, maize-wheat-soybean meal basal diet; 2) ME60, CON + 60 mg/kg multi-enzyme; 3) ME120, CON + 120 mg/kg multi-enzyme; 4) ME180, CON + 180 mg/kg multi-enzyme. Increasing concentration of multi-enzyme had positive linear effects on body weight gain and feed conversation ratio from days 7 to 21, 21 to 35, and 1 to 35 (P<0.05). Supplementation with multi-enzyme did not affect feed intake throughout the experiment (P>0.05). Additionally, the apparent total tract digestibility of dry matter and crude protein increased linearly with increased levels of multi-enzyme (P<0.05). These data indicated that supplementation with 180 mg/kg multi-enzyme was effective for improving growth performance and nutrient digestibility in broilers fed with maize-wheat-soybean meal-based diets. [ABSTRACT FROM AUTHOR]

A designed biocatalytic cascade system based on reverse enzymatic catalysis by formate dehydrogenase (EC 1.2.1.2), formaldehyde dehydrogenase (EC 1.2.1.46), and alcohol dehydrogenase (EC 1.1.1.1) can convert carbon dioxide (CO 2 ) to methanol (CH 3 OH) via formation of formic acid (CHOOH) and formaldehyde (CHOH) during equimolar cofactor oxidation of NADH to NAD + . This reaction is appealing because it represents a double gain: (1) reduction of CO 2 and (2) an alternative to fossil fuel based production of CH 3 OH. The present review evaluates the efficiency of different immobilized enzyme systems and reaction designs that have been explored for optimizing this sequential enzymatic conversion of CO 2 to CH 3 OH, including multilayer microcapsules, bead scaffolds, cationic nanofibers, and membrane systems. The recent progress within efficient cofactor regeneration, protein engineering of the enzymes for robustness, and advanced uses of membrane systems for enzyme reuse and product separation are assessed for large scale implementation of this biocatalytic reaction cascade. Industrial realization of enzymatic CO 2 to CH 3 OH conversion including the option for reaping of formaldehyde and formate during the reaction warrants innovative development. There is a particular need for development of i) better enzymes; ii) improved understanding of enzyme structure function aspects of reverse catalysis by dehydrogenases, iii) quantitative kinetic models of the enzymatic cascade reaction during simultaneous cofactor regeneration, iv) robust systems for regeneration of reducing equivalents. [ABSTRACT FROM AUTHOR]

To improve the efficiency of enzymatic saccharification for lignocellulose, an efficient crude multi-enzyme was produced by Trichoderma reesei using corncob, a low cost inducer. Expression of cbh1, bgl1, egl1, xyn1 and positive regulator xyr1 induced by corncob increased significantly compared to that by cellulose. After 120 h induction by corncob, enzymatic activities on filter, CMC, β-glucose and xylan increased 86.5, 46.9, 120.9 and 291.2% compared to those induced by cellulose, and the concentration of secreted protein increased by 120.8%. FPase:β-glucosidase and FPase:xylanase values in crude multi-enzyme I (ECI, induced by corncob) were higher than that in crude multi-enzyme II (ECII, induced by cellulose). Under the same hydrolysis conditions, the volume dosage of ECI was only half of ECII, but ECI still showed a maximum of 12.5 and 33.4% higher than ECII in the total reducing sugar and glucose yield in lignocellulose hydrolysis. Corncob could be a candidate for low cost production of multi-enzyme for efficient lignocellulose degradation, and this work could guide the genetic modification of T. reesei to obtain efficient multi-enzyme for lignocellulose hydrolysis. [ABSTRACT FROM AUTHOR]

Dual enzyme-responsive peptides were synthesized by masking the ɛ-amine of lysine with various enzyme substrates. Enzymatic cleavage of these sequences unmasked the ɛ-amine, allowing for further digestion by a second enzyme, which was monitored colorimetrically. This modular peptide design should provide substrates for a large combination of clinically relevant enzymes. [ABSTRACT FROM AUTHOR]

This study was designed to investigate the influence of supplementing multi-enzyme in feed on growth and survival rate of rainbow trout fish. The multi-enzyme were incorporated into a commercial pellet ration to constitute three experimental diets containing 500, 1000, 1500 gram kemin phytase multi-enzyme per one ton feed intake, while a fourth diet without the multi-enzyme supplement served as the control diet. The diets were offered to rainbow trout (150 g average weight) in quadruplicate pools (each treatment) for 40 days. Fish were sampled at 10, 20, 30 and 40 days after commencement of the feeding trial to determine feed conversion ratio (FCR), weight and length rate and survival rate. Results of our study indicated that effect of different supplementing multi-enzyme levels on feed conversion ratio (FCR), weight and length rate and survival rate, there was not any meaning relation in dose rates (p>0.05). According to affect of type and concentrates of enzyme on animal response, we can recommend that type or concentrates of consummated kemin- phytase multi-enzyme is not suitable for rainbow trout.

We developed a synthetic route for producing 3-amino-2-hydroxy acetophenone (3AHAP) from m-nitroacetophenone (3NAP) using an in vitro approach. Various reaction systems were evaluated, and a direct reaction method with crude enzyme and supersaturated substrates for optimal catalytic efficiency was chosen. The reaction system included three enzymes and was enhanced by adjusting enzyme molar ratios and optimizing ribosomal binding sites. We performed substrate docking and alanine scanning to identify key sites in the enzymes nitrobenzene nitroreductase (nbzA) and hydroxylaminobenzene mutase (habA). The optimal mutant was obtained through site-directed mutagenesis, and incorporated into the reaction system, resulting in increased product yield. After optimization, the yield of 3AHAP increased from 75 mg/L to 580 mg/L within 5 hours, the highest reported yield using biosynthesis. This work provides a promising strategy for the efficient and sustainable production of 3AHAP, which has critical applications in the chemical and pharmaceutical industries., (© 2023 Wiley-VCH GmbH.)

The major complications in fruit juice quality improvement are the presence of polysaccharides components in the form of disrupted fruit cell wall and cell materials. Hence, breakdown of cellulose along with pectin and starch is important for the juice processing. In this context, magnetic tri-enzyme nanobiocatalyst was prepared by simultaneously co-immobilizing three enzymes; α-amylase, pectinase and cellulase onto amino-functionalized magnetic nanoparticle by 60 mM glutaraldehyde concentration with 10 h cross-linking time for one pot juice clarification. The prepared nanobiocatalyst was characterized by FT-IR, SEM and XRD. The thermal (50–70 °C) and pH (3–6) stability studies indicated more than two folds increment in half-life and enhanced tolerance to lower pH. The immobilized enzymes retained up to 75% of residual activity even after eight consecutive cycles of reuse. Finally, the clarification of apple, grapes and pineapple juices using magnetic tri-enzyme showed 41%, 46% and 53% respective reduction in turbidity till 150 min treatment. [ABSTRACT FROM AUTHOR]

The objectives were to determine the standardized ileal digestibility (SID) of AA and NE value of cold-pressed soybean cake (CP-SBC), and the effect of extrusion or adding multi-enzyme to CP-SBC diet for growing pigs. Eight ileal-cannulated pigs (initial BW = 79.7 ± 3.97 kg) were fed 4 diets in a replicated 4 × 4 Latin square design to give 8 replicates per diet. Diets included a cornstarch-based diet with CP-SBC, extruded CP-SBC, and SBC plus multi-enzyme (1,200 U of xylanase, 150 U of glucanase, 500 U of cellulase, 60 U of mannanase, 700 U of invertase, 5,000 U of protease, and 12,000 U of amylase/kilogram of diet; Superzyme-CS, 0.5 g/kg); and a N-free diet. The CP-SBC was the sole source of protein in the CP-SBCcontaining diets. The ratio of cornstarch to sugar and soybean oil in CP-SBC-containing diets was identical to the N-free diet to allow calculation of energy digestibility of CP-SBC by the difference method. The evaluated CP-SBC had been produced by heating the soybean seed at 105°C for 60 min followed by pressing of the heated soybean seeds at less than 42°C (barrel temperature). On a DM basis, CP-SBC and extruded CP-SBC contained 47.8 and 47.1% CP, 15.6 and 10.5% ADF, 7.23 and 8.85% ether extract, 3.11 and 3.08% Lys, and 2.25 and 3.70 trypsin inhibitor units per mg, respectively. Extrusion increased (P < 0.001) the SID of AA for the CP-SBC by an average of 12%. Also, extrusion increased (P < 0.001) the NE value of the CP-SBC from 2,743 to 2,853 kcal/kg of DM. Supplementation of CP-SBC diet with the multi-enzyme increased (P < 0.05) the SID of Arg and Pro, and tended to increase (P < 0.1) the SID of Ile and Tyr. However, the multienzyme supplementation did not affect the NE value of CP-SBC. In conclusion, the CP-SBC evaluated in the present study could be an alternative source of AA and energy in swine diets, and its nutritive value can be increased by extrusion following cold-pressing. The multi-enzyme used in this study improved the digestibility of some AA, but had limited effect on energy digestibility and hence NE value of the CP-SBC. [ABSTRACT FROM AUTHOR]

This study sought to evaluate the effect of Alchornea cordifolia leaf meal (ACLM) inclusion and exogenous enzyme supplementation on broiler chickens. Two hundred and fifty-two (252) arbour acre two weeksold broiler chicken were selected from a larger flock that had been brooded on deep litter and raised on commercial diet. These birds were balanced for weight and thereafter distributed into 18 pens of 14 birds each. These pens were randomly allotted to six dietary treatment groups in such way that 3 replicates were fed on each experimental diet. For the starter and finisher phase, six experimental diets were formulated and designated as 0-E, 5-E and 10-E for the diets without enzyme and 0+E, 5+E and 10+E for diets with enzyme. Diet 0-E and 0+E serve as positive and negative control respectively. The experiment was carried out using a 2x3 factorial experiment comprising of 2 enzyme levels (0 and 0.35 g/kg) and 3 ACLM inclusion levels (0, 5 and 10%).The final weight (FW) and weight gain (WG) were significantly affected by enzyme supplementation (p<0.05) while ACLM inclusion levels caused significant (p<0.05) decrease of 15.13% and 11.61% in the total feed intake (TFI) and feed conversion ratio (FCR) respectively in the broiler chickens. Enzyme supplementation significantly (p<0.05) increased the slaughtering weight, drum stick and thigh by 4.90%, 5.06 and 5.33 % respectively, while the ACLM inclusion level reduced the dressing percentage, drum stick, wing weight, thigh, breast and back significantly (p<0.05), decreasing by 17.63%, 25.95%, 10.72%, 9.85%, 21.27% and 19.41% respectively, as the ACLM level increase ranged from 0 to 10 %. The internal organs of the broilers were not significantly affected (p>0.05) by the main factors and their interactions except for the liver and proventriculus that were affected by the level of inclusion of ACLM. Red blood cells (RBC) significantly increased (p<0.05) while mean cell haemoglobin (MCH), mean cell volume (MCV) and monocytes significantly (p<0.05) decreased with an increase in the level of ACLM inclusion ranging from 0 to 10%. Enzyme supplementation caused a significant (p<0.05) increase in cholesterol and SGPT levels and a significant (p<0.05) decrease in SGOT. ACLM also significantly (p<0.05) affected the serum parameters considered, except for total protein, albumin, globulin, creatinine, bilirubin and SGPT in a way in which the increase of the ACLM level ranging from 0 to 5% and then to 10 % in the broiler's diets promoted the declination in the cholesterol, urea, SGOT and glucose levels. Furthermore, the interaction of enzyme levels and ACLM has a significant (p<0.05) effect on cholesterol and SGPT levels. Basing on the data obtained in this study, ACLM is a potential useful feed material for feeding monogastrics. Therefore, it is suggested that 10% ACLM inclusion level could be used in broiler production. [ABSTRACT FROM AUTHOR]

The objectives were to determine the standardized ileal digestibility (SID) of amino acids (AA) and AMEn value of cold-pressed camelina cake (CPCC) and the effect of adding multi-enzyme to a corn-CPCC diet for broilers. The 600 male broiler chicks were divided into 40 groups and fed 5 diets in a completely randomized design (8 groups per diet) from d 15 to d 21 of age. A corn basal diet and the basal diet with 30% of it replaced by CPCC were used in a 2 × 2 factorial arrangement with or without multienzyme (2,800 U of cellulase, 1,800 U of pectinase, 400 U of mannanase, 50 U of galactanase, 1,000 U of xylanase, 600 U of glucanase, 2,500 U of amylase, and 200 U of protease/kilogram of diet; Superzyme OM, 1 g/kg). The fifth diet was N-free. The corn basal diet was fed to determine nutrient digestibility and retention for CPCC by substitution. The N-free diet was fed to estimate basal endogenous AA losses for determining SID of AA. Diets contained TiO2 as indigestible marker. On a DM basis, CPCC contained 39.8% CP, 38.3% neutral detergent fiber, 12.7% ether extract, 1.89% Lys, 0.70% Met, 1.56% Thr, and 0.45% Trp. The SID of Lys, Met, Thr, and Trp for CPCC were 76.5, 85.5, 72.8, and 84.1%, respectively. The AMEn value for CPCC was 1,671 kcal/kg of DM. Multi-enzyme supplementation increased (P < 0.05) the SID of Met and Thr and the AMEn value of the corn-CPCC-based diet by 1.4, 1.3, and 3.0%, respectively. The multi-enzyme increased (P = 0.026) the AMEn value of CPCC from 1,671 to 1,941 kcal/kg of DM. In conclusion, the CPCC evaluated in the present study can be included in poultry diets as a source of energy and AA. Multi-enzyme supplementation increased the AMEn value of CPCC for broilers. [ABSTRACT FROM AUTHOR]

Mariculture organic waste (MOW) is rich in organic matter, which is a potential energy resource for anaerobic digestion. In order to enhance the anaerobic fermentation, the MOW was hydrolyzed by multi-enzyme and thermophilic bacteria. It was advantageous for soluble chemical oxygen demand (SCOD) release at MOW concentrations of 6 and 10 g/L with multi-enzyme and thermophilic bacteria pretreatments. For multi-enzyme, the hydrolysis was not obvious at substrate concentrations of 1 and 3 g/L, and the protein and carbohydrate increased with hydrolysis time at substrate concentrations of 6 and 10 g/L. For thermophilic bacteria, the carbohydrate was first released at 2-4 h and then consumed, and the protein increased with hydrolysis time. The optimal enzyme hydrolysis for MOW was determined by measuring the changes of SCOD, protein, carbohydrate, ammonia and total phosphorus, and comparing with acid and alkaline pretreatments. [ABSTRACT FROM AUTHOR]

A 4 × 3 factorial experiment was designed to study the effects of a multi-enzyme (Avizyme 1502; combination of xylanase, α-amylase and protease) in three levels (0, 250, 500 mg/kg) on performance and carcass characteristics of broiler chickens fed corn-soybean meal diets with different levels of metabolizable energy (ME; 13.18; 12.34; 11.93 and 11.51 MJ/kg) from 1 to 49 days of age. Results showed that dietary treatments had no significant effect (P> 0.05) on feed intake at 10 days and 24 days but birds fed diets containing 13.18 MJ/kg ME with 500 mg/kg multi-enzyme consumed more feed than birds fed diets with 11.51 MJ/kg ME and 250 mg/kg multi-enzyme (P< 0.05) at 49 days. Average daily gain in starter (1–10 days) and finisher (25–49 days) periods was affected by interaction between ME and enzyme. No significant differences (P> 0.05) were observed for feed conversion ratio (FCR) in grower period, although dietary treatments affected FCR in starter and finisher phases. At 49 days of age, the best FCR obtained for birds fed diets containing 13.18 MJ/kg ME and 250 mg/kg multi-enzyme. The highest FCR was for birds fed a diet with 11.51 MJ/kg ME with 250 mg/kg multi-enzyme. Breast, thigh and liver were significantly affected by dietary treatments. A diet with 13.18 MJ/kg ME and 500 mg/kg multi-enzyme resulted in the highest breast and thigh yield (P< 0.05). It can be concluded that adding enzymes to corn-soy based diets allowed the reduction in the energy level of broiler diets without any negative effects on performance of broiler chickens. [ABSTRACT FROM PUBLISHER]

Organophosphate compounds are heavily used in agriculture and military activities, while non-organophosphate pesticides are mostly used in agriculture and home defense. Discriminative detection of such toxic compounds is very challenging and requires sophisticated and bulky instrumentation. Meanwhile, multi-enzyme biosensors may offer an effective solution to the problem and may become a versatile analytical tool for discriminative detection of different neurotoxins. In this study, we report for the first time a novel bi-enzyme biosensing system incorporating electrostatically interacted enzyme-armored MWCNT–OPH and MWCNT–AChE along with a set of cushioning bilayers consisting of MWCNT–polyethyleneimine and MWCNT–DNA on glassy carbon electrode for discriminative detection of organophosphorus (OP) and non-organophosphorus (non-OP) pesticides. LbL interfaces were characterized by surface plasmon resonance and electrochemical impedance spectroscopy, demonstrating stepwise assembly and electron conductivity studies. The detection limit was found to be ~0.5 for OP pesticide paraoxon and 1 μM for non-OP pesticide carbaryl, in a wide linear range. The biosensor performance was also validated using apple samples. Remarkable discriminative and straightforward detection between OP and non-OP neurotoxins was successfully achieved with cyclic voltammetry (CV) and UV–vis methods on the MWCNT–(PEI/DNA) 2 /OPH/AChE biosensor, showing great potential in large screening of OP and non-OP pesticides in practical applications. [ABSTRACT FROM AUTHOR]

The hydrolysis effect of waste sludge after multi-enzyme and thermophilic bacteria pretreatments is investigated using excitation–emission matrix (EEM) with fluorescence regional integration (FRI) in this study. The compositional characteristics of extracellular polymeric substances (EPS) and dissolved organic matters (DOM) were analyzed to evaluate the sludge disintegration. The EPS and cell wall in sludge were disrupted after hydrolysis which led to carbohydrate, protein and soluble chemical oxygen demand (SCOD) of DOM increasing in sludge supernatant. The bio-degradability level in the extracted fractions of EPS and DOM depending on the fluorescence zones was found after hydrolysis. The highest proportion of percent fluorescence response ( P i , n ) in EPS and DOM was soluble microbial by-product and humic acid-like organics. A significant increase of humic acid-like organics in DOM after thermophilic bacteria hydrolysis was obtained. The assessment of hydrolysis using EEM coupled with FRI provided a new insight toward the bio-utilization process of waste sludge. [ABSTRACT FROM AUTHOR]

Abstract: Globotriose is involved in numerous pathogenic processes and drug development strategies. Recent studies have demonstrated that globotriosylceramide could be used in colon cancer therapy and as a crucial indicator for susceptibility to HIV-1 infection. Therefore, the cost-effective and facile approaches for large-scale production of globotiose as potential drugs are highly required. Here, a multi-enzyme one-pot system containing a galactokinase (SpGalK, E.C.2.7.1.6), a UDP-glucose pyrophosphorylase (SpGalU, E.C.2.7.7.9), a α-1,4-galactosyltransferase (LgtC, E.C. 2.4.1.44) and a commercial inorganic pyrophosphatase (PPase, EC 3.6.1.1) was designed to achieve globotriose on preparative scales. This method exploits a cheaper initial substrate, galactose, for donor UDP-galactose production. More importantly, the substrate specificity of SpGalK and SpGalU is highly promiscuous and various UDP-galactose derivatives obtained could be used as the donor substrates for LgtC. This pointcut of rapid preparation of globotriose derivatives is proposed for the first time. Finally, three globotriose analogs were achieved by this one-pot multi-enzyme system in our study. [Copyright &y& Elsevier]

Two experiments were conducted to assess the effects of dietary supplementation of an exogenous multi-enzyme (EME) preparation to 35-to 65-d-old piglets on apparent total tract digestibility (ATTD), growth performance, digestive enzyme activities, and selected microbial populations in feces. In Exp.l, twenty eight 35-d-old piglets were randomly assigned to 7 dietary treatments (corn-soybean based diet supplemented with 0, 100, 150, 200, 250, 300, or 350 mg EME/kg) in a 14-d digestibility study. Piglets fed the diets supplemented with EME had greater ATTD of DM, CP, and GE (P = 0.001,0.005, and 0.009, respectively) than those fed the diet without EME supplementation, and those ATTD values increased linearly and quadratically (P < 0.001) as the levels of supplemented EME increased. In Exp. 2, two hundred 35-d-old weanling piglets were randomly allocated to 20 pens. The pens were then randomly assigned to 5 dietary treatments (corn-soybean based diet supplemented with 0, 100, 150, 250, or 350 mg EME/kg) with 4 pens per treatment in a 30-d feeding experiment. Piglets has ad libitum access to diets and water, and they were weighed at the beginning (35-d-old), middle (50-d-old), and end (65-d-old) of the experiment. Fecal samples were grabbed directly from the rectum and digesta samples from duodenum, jejunum, and ileum were taken at the end of the experiment for the analysis of selected bacteria populations and digestive-enzyme activities. The ADG and ADFI tended to be greater with the increasing levels of supplemented EME in both periods, whereas G:F was improved (P = 0.012 and 0.017) by EME in the period of 35 to 50 d of age and during the overall experimental period. Furthermore, inclusion of EME in diet increased the counts of Lactobacilli spp. and Bacillus subtilis spp., but reduced the populations of Salmonella spp. and Escherichia coli spp. in the feces. The EME supplementation also enhanced (P < 0.05) the activities of amylase, lipase, and protease in the small intestine. The growth performance-enhancing effects of EME appeared to be mediated by the age of the piglet and the dose of EME used. Supplementation of corn-soybean meal diets for 35- to 65-d-old piglets with EME has a potential to enhance gut health condition, increase nutrient digestion, and increase growth performance. [ABSTRACT FROM AUTHOR]

An experiment was conducted to evaluate the efficacy of multienzyme supplement (Natuzyme®) in diets containing wheat on the broilers performance. A total of one hundred twenty day old male broiler chicks (Arbor Acres) were randomly assigned to 6 treatments with a 2 3 factorial arrangement (0, 150 and 300 gram per kilogram wheat and 0 and 0.025 gram per kilogram enzyme). Chickens were fed with starter, grower and finisher diets during 1-21, 22- 35, 36-49 days of the age, respectively. Body weight gain, feed intake, and feed conversion ratio were determined weekly and productivity efficiency index was determined at the end of experiment. Litter moisture was evaluated at the age 21 days. The results indicated that although different levels of multi-enzyme and wheat had no significant effect on body weight gain, feed intake, feed conversion ratio, productivity efficiency index and litter moisture but caused partial improvement in these parameters. [ABSTRACT FROM AUTHOR]