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Bating is one of the processes in leather production. This process is generally performed under slightly alkaline conditions, but it can also be done under acidic conditions. Rhizopus oligosporus is categorized as a microorganism that can synthesize acidic enzymes optimally in acidic conditions. This study aimed to evaluate the effect of the bating process using acid protease from R. oligosporus on the leather qualities tanned by chromium sulfate. The study used a completely randomized design where crude enzyme activity (0, 2.5, 5, 7.5, and 10 U mL-1) was a single treatment as an experimental design. Data were analyzed using ANOVA and furtherly tested using Duncan. The results show that the protease synthesized from R. oligosporus is potential to be used as a bating agent. The treatment with 2.5 U mL-1 activity was the best treatment in this study to improve leather (wet blue) properties like shrinkage temperature, ash content, and chrome oxide content. The quality of wet blue produced was better than that produced with conventional treatments. It can be concluded that the enzyme activity of 2.5 U mL-1 could be used as a bating-agent alternative in the acid bating stage.

The purpose of this research was to determine the dietary effect of red oncom ( Neurospora sitophila spp ) and black oncom ( Rhizopus oligosporus ) on performance and intestinal properties of broiler in different levels. The materials used for this research were 100 unsex broiler (20 days old) with Lohmann strain. Method was used in this experiment was Completely Randomize Design with 5 treatments and 4 replications. The treatment were as follows, T0: basal diet without additive, T1: basal diet + (0.25% red oncom ( Neurospora sitophila spp ) and 0.25% black oncom ( Rhizopus oligosporus ), P2: basal diet + (0.5% red oncom ( Neurospora sitophila spp ) and 0.5% black oncom ( Rhizopus oligosporus ), T3: basal diet + (0.5% red oncom ( Neurospora sitophila spp ) and 0.25% black oncom ( Rhizopus oligosporus ), and T4: basal diet + (0.75% red oncom ( Neurospora sitophila spp ) and 0.5% black oncom ( Rhizopus oligosporus ). The result showed that red oncom ( Neurospora sitophila spp ) and black oncom ( Rhizopus oligosporus ) has significantly enhanced ( p

empeh is an Indonesian traditional fermented food with rich nutrition and bioactive components. Rhizopus sp,especially Rhizopus oligosporus), lactic acid bacteria (Lactobacillus sp.), and yeast aremicroorganisms involved in Tempeh fermentation. An interesting offer of Bioinformatics (in silico method) as a supporting tool in molecular biology studies has emerged, such as in protein cleavage. This study utilized PeptideCutter application on ExPASyBioinformatics portal (https://web.expasy.org/peptide_cutter/) to cleave soy proteins glycinin G1, G2, G3, G4,G5,-conglycinin-chain,andchainusingavailable enzymes in the applicationwith twosimulations. Simulation I was conducted using enzyme complex produced by Lactobacillus sp.and Rhizopus oligosporus, whilesimulation II was used enzyme complex produced by Lactobacillus sp., Rhizopus oligosporus, and Klebsiella pneumonia. Simulation I was conducted using enzyme complex produced by Lactobacillus sp. and Rhizopus oligosporus, whilesimulation II was used enzyme complex produced by Lactobacillus sp., Rhizopus oligosporus, and Klebsiella pneumoniae. A total of 58 peptides was found from the simulation I and higher than simulation I (41 peptides). The bioactive peptides by the cleavages using PeptideCuttertool were dominated with dipeptides and only three peptides were in the form of tripeptides, namely Leu-Leu-Phe(glycininG1), Val-Val-Phe(glycininG5), and Arg-His-Lys(-conglycinin-chain). Bioactive peptides with antihypertensive and antidiabetic properties were mostly found in this in silicomethod of soybean cleavage.

Adlay is a nutritious grain that has the potential as an alternative food because it has a high protein and fat content of 14.10% and 7.90%, respectively. The use of Adlay as flour still has a weakness, namely the functional properties of Adlay flour such as low swelling power. One way to improve the characteristics of Adlay flour is by fermentation using Rhizopus oligosporus. The fermentation duration is the time that allows changes in the characteristics of the flour due to the fermentation. This study aims to determine the effect of differences in fermentation duration using Rhizopus oligosporus on the physical, chemical, and functional characteristics of Adlay flour produced from fermented Adlay seeds. This study used one factor, namely the lengths of fermentation for 0, 12, 24, 30, 36 and 48 hours. The analysis in this study included whiteness, yield, pH, water content, ash content, protein content, fat content, carbohydrate content, swelling power and solubility, oil holding capacity (OHC) and water holding capacity (WHC). The results have shown that the longer the fermentation duration the lower rate of whiteness, pH, yield, water content, fat content, and carbohydrate content will become, while some functional properties of Adlay flour become better with increasing values of ash content, protein content, swelling power, solubility, Oil Holding Capacity (OHC), and Water Holding Capacity (WHC).

Peanut protein concentrates (PPCs) were subjected to hydrolysis by crude protease extract (CPE) obtained from three fungi viz; Rhizopus oligosporus, Trichoderma reesei, and Aspergillus oryzae and the effect on structural, functional and in-vitro protein digestibility (IVPD) properties were studied. Particle size was found significantly (p ≤ 0.05) lower in hydrolyzed samples than un-treated samples. Fourier transform infrared spectroscopy (FTIR) spectrum of hydrolyzed samples displayed intense absorbance peaks in the wavelength ranging from 1500 to 2600 cm(−1). Peanut protein concentrates hydrolyzed by CPE from R. oligosporus showed higher surface hydrophobicity (564.18). Total sulfhydryl content was found lower in all the hydrolyzed samples whereas, reverse trend was observed for exposed sulfhydryl content. The structural changes simultaneously affected the functional and IVPD attributes of hydrolyzed PPCs. In comparison to the PPCs hydrolysed using crude extracts from T. reesei and R. oligosporus, PPCs hydrolysed by A, oryzae showed higher solubility, water and oil binding capacity, foaming capacity and foam stability. Higher IVPD values of 86.70% was also found in PPCs hydrolyzed with CPE of A. oryzae. The study established that CPE hydrolysis of PPCs has potential for scale-up studies and may serve as a cost effective alternative to protein hydrolysis with pure enzymes.

Made available in DSpace on 2022-05-01T06:02:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Purpose: This work aimed the food waste (FW) valorization by the production of fungal enzymes in solid-state fermentation (SSF) and a hydrolysate with the potential to obtain products with high added value. Methods: An original bioprocess based on Rhizopus oligosporus cultured by SSF using medium consisting of FW, sugarcane bagasse (SCB), wheat bran (WB), and corn steep liquor (CSL) utilizing a cell-substrate recycling system was implemented. Results: The highest outcomes were achieved using FW as the main substrate and the strategy of one cell recycling round. The best condition for amylase production (260.9 U/g) was the blend FW 50%, SCB 10%, WB 40% supplemented with a salt solution. For the highest protease level (665.5 U/g) the same mixture was supplemented by 20% CSL. Hydrolysis of FW with the enzymatic extract from the best specified blend produced 47.10 g/L of reducing sugars. Conclusion: This bioprocess showed to be cost-effective, technically not demanding, and could be scaled up successfully for commercial goals. Graphical Abstract: [Figure not available: see fulltext.] Bioprocess Unit Department of Biotechnology Bioenergy Research Institute (IPBEN) Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP), Campus Assis, Avenida Dom Antônio, 2100, SP Escola de Artes Ciências e Humanidades Universidade de São Paulo, Rua Arlindo Bettio, 1000, Vila Guaraciaba, SP Bioprocess Unit Department of Biotechnology Bioenergy Research Institute (IPBEN) Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP), Campus Assis, Avenida Dom Antônio, 2100, SP CNPq: 141472/2015-4 FAPESP: 2014/241881 FAPESP: 2017/029097 FAPESP: 2018/045076

Berdasarkan kajian-kajian teori, penelitian, dan fakta yang ada , ternyata tempe merupakan jenis makanan yang berdampak awet muda bagi penggemarnya. Dengan catatan, tempe yang dikonsumsi bermutu baik dan pola konsumsinya teratur (minimal 100 gram setiap hari). Dalam buku “Longevity Through The Organic Lifestyle”, yang ditulis oleh Henry Chang, dan diterbitkan tahun 1999 dijelaskan, tempe merupakan makanan hasil fermentasi, termasuk salah satu makanan untuk awet muda, sangat baik dikonsumsi oleh siapa saja, terutama yang sudah berusia lanjut.

This study aimed to investigate and optimize the quality of tomato ‘Soksungjang’. Tomato ‘Soksungjang’ was prepared using 10% tomato paste with the addition of Rhizopus oligosporus Koji. Various quality characteristics were determined including proximate composition, physicochemical properties, free sugar and organic acid contents, free amino acid content, total microbial count, enzyme activity, and the presence of volatile compounds. The results showed that the free sugar content of tomato ‘Soksungjang’ was 14.81 mg%. The individual sugars identified were glucose, galactose, fructose, sucrose, maltose, and lactose. The total organic acid content was 162.79 mg% and the identified individual organic acids were citric acid, lactic acid, acetic acid, malic acid, formic acid, and succinic acid. The total free amino acid content was 1,440 mg%, including major amino acids, such as glutamic acid, lysine, and arginine. The α-amylase activity of the tomato ‘Soksungjang’ was 74.42 unit/mL, whereas lipase and neutral protease activities were 93.59 and 35.79 unit/mL, respectively. Total 30 compounds including alcohols, esters, ketones, and aldehydes were detected in tomato ‘Soksungjang’. Ethyl alcohol, 3-methyl-1-butanol, and ethyl acetate were among the major volatile compounds.

The present study evaluated the detoxification and nutritional enrichment potentials of the fermented cassava waste pulp obtained through different processing methods of sweet (S) and bitter (B) cassava varieties by fermentation with Rhizopus oligosporus and Aspergillus niger. Four batches [two with both bark and cortex of the tuber removed (S- and B-), two with bark only removed (S+ and B+)] of cassava waste pulp (CWP) were produced from each of S and B varieties. CWP from the four treatments were then subjected to Rhizopus oligosporus, Aspergillus niger, and natural fermentation. Cyanide (mg HCN/kg), phytate (mg/100g), tannin (mg/100g), pH, crude protein (N X6.25) (%), ash (%), Fe, P, K and Ca (mg/100g) content of the unfermented and fermented CWP were determined using standard procedures. The range of the reduction (%) in cyanide content for Rhizopus oligosporus fermented CWP (RCWP), Aspergillus niger fermented CWP (ACWP), and naturally fermented CWP (NCWP) were 86.24 – 91.95, 79.68 – 84.34 and 72.96 – 78.58 while those for phytate were 58.82 – 71.20, 29.98 – 39.26 and 36.16 – 40.32 respectively. Fermentation had no significant (p>0.05) effect on the tannin content while the pH range of NCWP, RCWP and ACWP were 7.20 – 7.50, 5.41 – 6.40, and 3.83 – 4.21, respectively. The range of the increase (%) in the protein content for NCWP, RCWP, and ACWP was 346 – 375 , 671 – 728 and 508 – 542 while those of the ash content were 7.16 – 7.91, 6.72 – 6.84, and 5.42 – 5.63 respectively. Of all the examined minerals, only calcium increased by 5.68 – 26.89, 49.96 - 70.43, and 59.71 – 98.87 % in NCWP, RCWP, and ACWP respectively. Results from the present study are indicating that controlled fermentation offers greater benefit over natural fermentation and that the cassava variety and not the peeling method will determine the extent of detoxification and nutrient enrichment.

Generally, the microorganism involved in soybean fermentation for the production of tempeh is Rhizopus oligosporus. However, Saccharomyces cerevisiae, a type of β-glucan-producing yeast, is known to be present and grow in the fermentation process. This study was aimed at investigating yeast and fungal growth dynamics, β-glucan formation, and antibacterial activity against Escherichia coli during the fermentation after adding S. cerevisiae as an inoculum. The Randomized Complete Block Design (RCBD) was applied with two treatments and three repetitions. Three types of starter culture were S. cerevisiae, R. oligosporus, and the combination of both. The second treatment was fermentation time at room temperature ( 30 ± 2 ° C ) for 0, 8, 16, 24, 32, and 40 hours. The dynamics were observed every eight hours. The obtained data were tested using Tukey’s Honestly Significant Difference (HSD) test. The results indicated that yeast grew during this process from a single S. cerevisiae culture and a mixture of R. oligosporus and S. cerevisiae, but not from R. oligosporus alone. The yeast grew during and until the end of fermentation and decreased after 32 hours in the mixed cultures. The β-glucan formed in tempeh with all types of inoculum, but the antimicrobial activity against E. coli increased with fermentation time and was significantly different between treatments. The highest β-glucan content and antibacterial activity of tempeh are from the mixed culture. In conclusion, the addition of S. cerevisiae and R. oligosporus in soybean fermentation produced tempeh with the highest β-glucan content and antibacterial activity against E. coli. The presence of β-glucans suggests higher health benefits of tempeh.

The aim of this study was to determine the effect of both the Solid State Fermentation (SSF) technique and the use of Rhizopus oligosporus on the physicochemical changes of fermented de-husked rice flour. Three varieties of de-husked rice, i.e., Mentik Wangi Susu (non-pigmented), red Cempo Merah, and black Jowo Melik (pigmented) were fermented using Rhizopus oligosporus. Fermentation was performed at room temperature with a fermentation time of 0, 24, 48, and 72 hours. The analyzed parameters were proximate composition, bio-active compounds, and pasting profile. The results showed an increase in flour pasting profile, ash, protein, and fat content after the fermentation. The total availability of the total phenolic content (TPC) and antioxidant capacity were also increased. The highest TPC (0.37 mg GAE/g) and antioxidant capacity (1.43 mg TEAC/g) were obtained in the Jowo Melik variety at 72 hours of fermentation. In contrast, anthocyanin and carbohydrate contents decreased as fermentation time increased. The highest anthocyanin content of 0.53 mg/g (after 24-hour fermentation) was obtained in the Jowo Melik variety. In conclusion, 72-hour-fermented black rice flour (Jowo Melik) has a higher potential to be developed as a functional food.

Antibacterial activities against Staphylococcus aureus and Bacillus subtilis were found in an ethanol fraction of tempe, an Indonesian fermented soybean produced using Rhizopus oligosporus. The ethanol fraction contained free fatty acids, monoglycerides, and fatty acid ethyl esters. Among these substances, linoleic acid and α-linolenic acid exhibited antibacterial activities against S. aureus and B. subtilis, whereas 1-monolinolenin and 2-monolinolenin exhibited antibacterial activity against B. subtilis. The other free fatty acids, 1-monoolein, monolinoleins, ethyl linoleate, and ethyl linolenate did not exhibit bactericidal activities. These results revealed that R. oligosporus produced the long-chain polyunsaturated fatty acids and monolinolenins as antibacterial substances against the Gram-positive bacteria during the fungal growth and fermentation of heat-processed soybean.

This study was performed to verify the possibility of manufacturing Rhizopus oligosporus ‘Koji’, which is cultured using a range of different substrates. The growth characteristics of R. oligosporus were therefore investigated for the preparation of R. oligosporus ‘Koji’, and the optimal incubation conditions were determined to be 30 °C for 24 h, with an optimal NaCl concentration were 5%. Based on these results, R. oligosporus was incubated using three substrates, namely beans, rice and a mixture of beans and rice, for 6 d, and the enzyme activities and volatile compounds of the fermentation products were investigated. On the fourth day of incubation, the α-amylase activities of the rice and the mixed substrates were 155 and 42 unit/mL, respectively, while on the fifth day of incubation, the neutral protease activities of the beans and the mixed substrate were 14 and 15 unit/mL, respectively. In terms of the lipase activity, the mixed substrate resulted in a higher activity than the other substrates. Various volatile flavor compounds, including alcohols, ketones, esters, aldehydes, and pyrazines, were identified in R. oligosporus ‘Koji’ cultured using the mixed substrate.

Purpose The purpose of this paper is to understand the effect of hydrolysis by pepsin and pancreatin on the angiotensin-I-converting enzyme (ACE) inhibitory activity of bioactive peptide from pigeon pea tempe and the absorption of pigeon pea tempe peptide by using the everted gut sac method. Design/methodology/approach The tempe was prepared by inoculating Raprima (Rhizopus oligosporus) on hulled-cooked pigeon pea for 48 h. The extraction was performed using the ultrasonic method at 40 kHz frequency and 100% ultrasonic power for 10 min. The extracted protein was placed in simulated gastrointestinal digestion using consecutive pepsin–pancreatin for 240 min. The hydrolysates were fractionated using a dialysis tube, and its absorption was assessed using the everted Sprague–Dawley rat gut sac. Findings The tempe protein from the hydrolyzed pigeon pea exhibited higher ACE inhibitory (71.53%) activity than that from the boiled pigeon pea (53.04%) (p = 0.028). The bioactive peptide of the digested pigeon pea tempe consisted of low-molecular-weight peptides (p = 0.021). Bioactive peptides from pigeon pea tempe were absorbed well in the small intestine, mainly in the jejunum. The activity of the absorbed peptides did not change considerably. Originality/value The activity of bioactive peptide of pigeon pea tempe was comparatively stable during digestion. It exhibited activity even after absorption in the small intestine. Thus, pigeon pea tempe can serve as an antihypertensive peptide source and alternative food for maintaining/reducing blood pressure.

The aims of this study were to determine the changes in physicochemical characteristics, microbiological safety and sensory acceptability of coconut dregs during fermentation using Rhizopus oligosporus. Fermentation of coconut dregs was done using Rhizopus oligosporus as a starter culture for three days (fresh coconut dregs was used as control). The control sample is fresh dregs without any process while other samples will through fermentation. The physicochemical analyses included internal temperature, pH, total soluble solids (TSS), water activity (aw), colour, proximate analysis and texture profile. Microbiological analyses included total plate count (TPC), Escherichia coli, Salmonella spp., Pseudomonas aeruginosa, Bacillus cereus, Staphylococcus aureus and LAB. Sensory acceptability was also carried out. The results showed that internal temperatures and TSS were increased and the pH decreased throughout the fermentation. The aw showed no changes. The colour of fermented coconut dregs had a higher percentage in lightness compared to yellowness and redness which showed that the colour of fermented dregs still maintains the white colour. A gradual increased in hardness, springiness, cohesiveness, chewiness and resilience was observed. Moisture content, crude fat and protein increased while ash and crude fibre decreased but no changes in carbohydrate throughout the fermentation. The results for microbial count showed the reducing number of S. aureus and B. cereus while increasing number of TPC and LAB. Interestingly, E. coli, Salmonella spp. and P. aeruginosa was not detected in fermented and non-fermented coconut dregs. The non-fermented coconut dregs were preferred more by the panellists as it obtained the highest score in term of colour, odour, texture and overall acceptability on sensory attributes evaluation.

Minyak kelapa murni merupakan bahan makanan yang banyak digunakan sebagai bahan konsumsi dan bahan tambahan untuk proses industri. Minyak kelapa murni diproduksi dari bahan baku krim santan yang merupakan hasil pelindihan antara air dengan buah kelapa. Pemecahan emulsi pada santan menggunakan teknik ekstraksi kering (dry extraction) dan teknik ekstraksi basah (wet extraction). Penelitian yang dilakukan menggunakan proses ekstraksi basah, yaitu secara fermentasi menggunakan inokulum padat berupa mikroba atau ragi yang dikembangkan pada media padat. Proses tersebut dipilih untuk menyesuaikan dengan situasi masyarakat daerah di Indonesia yang banyak menghasilkan kelapa serta menyesuaikan dengan keberadaan ragi tempe yang telah memasyarakat. Tujuan penelitian ini adalah untuk mendapatkan minyak kelapa dari bahan baku santan (coconut milk) dengan menggunakan inokulum ragi tempe yang mengandung Rhizopus oligosporus. Hasil minyak kelapa murni akan dianalisa parameternya berdasarkan Standar Nasional Indonesia. Dari data percobaan di peroleh hasil analisa produk minyak kelapa murni yang paling optimum sesuai dengan SNI adalah percobaan pada variasi jumlah ragi tempe 0.5 gram dengan jumlah minyak yang dihasilkan 91.35 gram dan nilai kekeruhan 1,8 NTU. Hasil analisis kimia dan fisik: Indeks bias: 1,451, Kadar air: 0,3%, Kadar kotoran: 0,023%, Kadar bebas asam lemak: 0,96%, Kadar minyak Pelikan: 1,82%, angka saponifikasi: 262,969 mgKOH / g lemak. Virgin coconut oil is a food ingredient that is widely used as a consumption material and as an additive for industrial processes. The oil produced from coconut milk cream which was the result of leaching between water and coconut. The emulsion in coconut milk then broken by dry extraction method and wet extraction method. In this research the method carried out by a wet extraction process, which used fermentation using microbes or yeast that was developed on solid media. This process was chosen to adapt to the Indonesian people who produced ton coconut each year and in the same time, the tempeh yeast that has been popular in Indonesia. The purpose of this study was to obtain virgin coconut oil from coconut milk using yeast inoculum containing Rhizopus oligosporus. The parameters of virgin coconut oil analyzed based on the Indonesian National Standard (SNI). The experimental data obtained from the results of the most optimal analysis of virgin coconut oil in accordance with SNI experiments on the variation of the amount of tempeh yeast 0.5 grams with the amount of oil produced 91.35 grams and turbidity value 1.8 NTU. Chemical and physical analysis results: Refractive index: 1,451, Air content: 0,3%, Dirt content: 0,023%, Free fatty acid level: 0,96%, Pelicans oil content: 1,82%, saponification rate: 262,969 mg KOH /g fat.

This article presents an innovative method of bio-mediated soil improvement for increasing the shear strength of loose sand. The improvement is realized by mixing the loose sand with the inoculum of Rhizopus oligosporus, a kind of fungus widely used in food industry for making Indonesian tempeh. The objective of this article is to investigate the performance and mechanism of mixing tempeh inoculum as a binding agent of loose sand particles. The inoculum dosage, water content of loose sand, and curing time were examined for identifying the increment of unconfined compressive strength (qu) of the samples. The results showed that qu of the treated samples increased when the inoculum dosage was elevated. It shows that 5.24% inoculum could yield 68 kPa of qu, and 5% water content and 3 d curing time produced the maximum qu. Moreover, the mechanism of hypha and mycelium in binding the soil particles was clearly observed using a digital microscope and scanning electron microscope. Keywords: Bio-mediated soil improvement, Loose sand, Rhizopus oligosporus

Most studies have found that vitamin B12 in tempeh is produced by contaminating bacteria specifically, Klebsiella sp. and Citrobacter freundii, during fungal fermentation. This study is to evaluate the effect of starter culture on the vitamin B12 and isoflavone aglicone content of soybean fermentation for tempeh production. In this study, soybeans were washed, soaked in water overnight, dehulled and sterilized by boiling at 100 ℃ for 30 min. Three starter cultures (103 CFU g-1) namely Rhizopus oligosporus, Klebsiella sp. and Saccharomyces cerevisiae were then inoculated as follows, soybeans + R. oligosporus + Klebsiella sp. (SRK), soybeans + R. oligosporus + S. cerevisiae (SRSc), soybeans + R. oligosporus + S. cerevisiae + Klebsiella sp. (SRScK), and Soy + R. oligosporus (SR) and soybeans + Klebsiella sp (SK). Inoculated soybeans were then incubated at 30 ±2 ℃ for 40 hours (tempeh-style). The growth of Klebsiella sp., S. cerevisiae, R. oligosporus and the production of vitamin B12 as well as isoflavone aglicones were observed. The results showed the highest vitamin B12 3.15 mg 100 g-1 was found in tempeh SRSc, followed by 2.88 mg 100 g-1 and 1.64 mg 100 g-1 in tempeh SR and SRScK, respectively. In addition, vitamin B12 in tempeh SRK was the lowest (0.81 mg 100 g-1). All the starter cultures were able to hydrolyze daidzin and genistin, but the amount of daidzein and genistein was tripled and doubled, respectively when Klebsiella sp was inoculated to the soybean fermentation. The study suggested that S. cerevisiae contributes to the production of vitamin B12, while Klebsiella sp contributes to production of daidzein and genistein in soybean fermentation for tempeh production.

This study investigated the formation of angiotensin-converting enzyme (ACE) inhibitory peptides from koro kratok beans tempe during gastrointestinal digestion. The absorption of bioactive peptides was also investigated in this study. Koro kratok was fermented by commercial culture including Rhizopus oligosporus for 48 h. Gastrointestinal digestion was simulated sequentially by hydrolysis of tempe protein extract with pepsin and pancreatin for 240 min. The peptide content, degree of hydrolysis, molecular weight distribution, and ACE inhibitory activity were analyzed. The absorption of ACE inhibitory peptides was evaluated using the inverted gut sac of Sprague Dawley rats. Results showed that some amino acids, such as Arg, Lys, Asp, Glu, Phe, and Leu, were predominantly found in tempe. After the hydrolysis process, cooked tempe exhibited the highest ACE inhibitory activity (90.05%). Although the ACE inhibitory activity of nonfermented koro kratok was lower than that of tempe, the increase in its inhibitory activity was too large (23.03%). The ACE inhibitory peptides from tempe showed a predominance of peptides with a molecular weight of

Extracts were extricated from raw okara and okara fermented with Rhizopus oligosporus using a clean, green protocol; water was used as the extraction solvent and coupled with ultrasound assistance for enhanced extraction. In vitro anti-oxidant analyses for antioxidant potential and capacity, superoxide scavenging activity, and nitric oxide scavenging activity validated that fermented okara yielded superior bioactive performance compared to raw okara. Fermented okara extracts showed no toxicity to erythrocytes and successfully prevented induced haemolysis. After 48 h incubation at the highest tested concentration (100 mg/mL), fermented okara extracts could inhibit HepG2 cells by 48.47 ± 5.28%, which was significantly different from their effects on NIH 3T3 cells. Gas chromatography–mass spectrometry characterization of extracts validated amino acids to be the chief fraction responsible for the detected bioactivity of the fermented okara extract. The results derived in this study open up the possibility that biofermented okara extract may be a potential novel sustainable nutraceutical.

Rice-derived bioactive compounds are now widely used as active ingredients in health foods, pharmaceutical and cosmeceutical products and as food additives. Polyphenolic compounds, gamma oryzanol, and vitamin E are among the bioactive constituents in rice that have important antioxidant, anti-aging, and anti-pigmentation properties among others. In this present study, broken rice was subjected to solid-state fermentation for 20 days by using two types of fungi, Aspergillus niger and Rhizopus oligosporus. Cosmeceutical-related functionalities of the fermented broken rice extracts were tested by evaluating antioxidant activities as well as tyrosinase and elastase inhibition activities. Total phenolic content and total flavonoid content of fermented broken rice extracts were also measured. Fermentation caused a substantial increase in the total phenolic and flavonoid content of broken rice. Ferric-reducing antioxidant activity was the highest (18.47 mM FE(II)/g sample) in broken rice fermented with R. oligosporus on day 20. Fermentation of broken rice with the same fungus resulted in higher DPPH radical-scavenging antioxidant activity than A. niger, with the highest activity recorded was 94.17%. Broken rice fermented with R. oligosporus for 20 days had the highest tyrosinase and elastase inhibition activities with the value of 46% and 51.9%, respectively. The results demonstrate that the fermentation process increased the biological activities and the numbers of bioactive compounds for potential use in the cosmeceutical industry. This undervalued by-product of the rice milling industry can effectively be used for the production of high value products through fungal solid-state fermentation. Keywords: Broken rice, Cosmeceuticals, Solid-state fermentation, Antioxidant

Soy isoflavones possess antioxidative, anti-inflammatory, anti-diabetic and phytoestrogenic properties. Soybean residue contains a fair amount of nutrients such as glycosylated isoflavones, minerals and dietary fibers, and is a substantial waste product produced from soymilk and tofu manufacturing. A solid-state fermentation of soybean residue by Rhizopus oligosporus or co-inoculated with Lactiplantibacillus plantarum improves the availability of isoflavones and GABA content which is attributed to ameliorated hyperglycemic symptoms in STZ-induced hyperglycemic mice. The effortless solid-state fermentation with present microbial manipulation supports an anti-hyperglycemia value-added application of soybean residue for functional food development. Background: Due to an awareness of the food crisis and with a rapidly rising prevalence of diabetes, recycling the substantial fibrous soybean residue disposed from soy industries has received consideration. Methods: Lactiplantibacillus plantarum was previously screened for active glutamate decarboxylase, and β-glucosidase activities were adopted for the fermenting of soybean residue using a traditional tempeh solid-state fermenting process with fungal Rhizopus oligosporus. Fermented soybean residue was chemically analyzed and functionally assessed in in vitro and in vivo hyperglycemic conditions. Results: A 48 h longer solid-state fermentation of the soybean residue co-inoculated with R. oligosporus and L. plantarum showed improved contents of isoflavone aglycones and GABA which were attributed to augmented antioxidative capacity, lowered ROS level, improved blood biochemistry, and better blood glucose homeostasis in STZ-induced hyperglycemic mice. Conclusion: The advantages of a food industrial effortless fermentation process, and a health nutritional endorsing anti-hyperglycemic value-added property offer a practical alternative in recycled soybean residue.

The aim of the present study was to evaluate the use of supercritical CO2 combined with cosolvent for the recovery of bioactive compounds of soybean fermented with Rhizopus oligosporus NRRL 2710. Soxhlet extractions using seven different organic solvents (n-hexane, petroleum ether, ethyl acetate, acetone, ethanol, methanol, and water) were initially performed for comparative purposes. The extracts obtained were characterized by physicochemical, antioxidant, total phenolic, and oxidative proprieties. For the Soxhlet extractions, the highest and lowest yields obtained were 45.24% and 15.56%, using methanol and hexane, respectively. The extraction using supercritical CO2 combined with ethanol as a static modifier (scCO2 + EtOH) presented, at a high pressure (25 MPa) and temperature (80 °C), a phenolic compound content of 1391.9 μg GAE g−1 and scavenging of 0.17 g, reaching a 42.87% yield. The extracts obtained by sCO2 + EtOH were characterized by high contents of essential fatty acids (linoleic acid and oleic acid) and bioactive compounds (gallic acid, trans-cinnamic acid, daidzein, and genistein). These extracts also showed a great potential for inhibiting hyaluronidase enzymes (i.e., anti-inflammatory activity). Thermogravimetric analyses of the samples showed similar profiles, with oil degradation values in the range from 145 to 540 °C, indicating progressive oil decomposition with a mass loss ranging from 93 to 98.7%. In summary, this study demonstrated the flexibility of scCO2 + EtOH as a green technology that can be used to obtain high-value-added products from fermented soybean.

PurposeThe purpose of this study was to evaluate thein vitroandin vivoeffects of three varieties of Indonesian fermented rice bran (RB) (Inpari 6, Inpari 30 and Inpara 1).Design/methodology/approachThree types of RB were fermented usingRhizopus oligosporus. The total phenolic content (TPC) was determined using the Folin–Ciocalteau method, and antioxidant activity was analyzed by measuring the 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity (RSA). Forin vivoanalyses, one week after acclimatization, stroke-prone spontaneously hypertensive rats (n= 4, 12 weeks of age) were divided into two groups and orally administered fermented RB (40 mg/kg body weight) or distilled water as a control after 16 h of fasting. Blood pressure (BP) was measured before and 2, 4 and 6 h after administration with a BP monitor without warming. Six days later, the rats were subject to the same procedure and sacrificed after 6 h of oral administration. Blood was collected and the plasma was separated to measure nitric oxide, glucose and insulin levels.FindingsThe highest TPC and RSA of fermented RB were obtained from Inpari 30 after incubation for 72 h (260.33 ± 0.39 mg GAE/100 g dry basis and 83.71 ± 0.61 per cent), respectively. Furthermore, single oral administration of fermented RB improved BP (p< 0.05) and glucose metabolism after 6 h of administration (p< 0.05).Originality/valueThis is the first study to evaluate the effects of fermented RB on improving high BP and glucose profiles by using a spontaneously hypertensive animal model.

Solid-state fermentation of soybeans by Rhizopus oligosporus RT-3 and Actinomucor elegans DCY-1 can be used as a natural enzyme pool for the improvement of dough and bread quality. Rheological analysis indicated that addition of tempeh flour (TF) increased G′ and G’’ moduli of dough. TF bread showed the largest specific volume (4.32 mL/g) and lowest hardness (113.64 g) on the first day. Bread substituted with TF exhibited less moisture loss in crumb (1.2 g/100 g) during the seven days storage. Molecular mobility and starch retrogradation of bread was measured by using low-field nuclear magnetic resonance (LF-NMR). Three different proton fractions, peaking at ∼0.16–0.24 ms (population A), ∼2.33–5.10 ms (population B) and ∼41.93–65.79 ms (population C), were detected by LF-NMR. Smaller changes in proton distributions (population A and B) of TF bread indicated that addition of tempeh flour hindered the shift of protons and the formation of amylopectin crystals during storage. TF bread was found to be sensorially acceptable and the highest scores for texture, aroma and volume were obtained. These results indicate that TF has great potential use for the improvement of bread quality and control of bread staling.

The use of protein hydrolysates (PHs) obtained by the enzymatic hydrolysis of underutilized proteins as active ingredients may be a promising strategy in the development of bioactive packaging. The effects of the incorporation of hydrolysed cottonseed proteins into alginate films were investigated in terms of their physical, chemical, barrier, optical, antioxidant and antimicrobial properties and the release of peptides in two different alginate film food simulants. PH incorporation did not affect the moisture content, biodegradability, solubility or oil barrier properties of the films but did increase the thickness and water vapor permeability. The increase in the PH concentration increased the barrier properties to visible light, and the film colour became darker, reddish and yellowish. The total phenolic content and the antioxidant activity (as tested by the DPPH, FRAP and ABTS methods) also increased. The PH films showed an inhibitory effect against Staphylococcus aureus, Colletotrichum gloeosporioides and Rhizopus oligosporus but not against Escherichia coli. In migration tests in aqueous media, the active films released more than 60% of their peptides in 30 min. Meanwhile, there was a controlled and gradual diffusion of the compounds embedded in the film when fatty foods were simulated. The results showed that alginate films with PHs show promise as active packaging for the preservation of fatty foods.

In this study, the bioaccessibility of D-chiro-inositol (DCI) from water biscuits formulated from raw and roasted buckwheat flours originating from common buckwheat after liquid-state fermentation (LSF) by select lactic acid bacteria (LAB) and fungi Rhizopus oligosporus 2740 was studied. The LAB-dependent variation in DCI content in fermented buckwheat flours was noted. LSF by L. salivarius AWH and R. oligosporus 2740 significantly enhanced the DCI content, whereas most of the applied LAB reduced DCI in the fermented flours. Baking at 220 °C for 30 min significantly enhanced the DCI level; however, no correlation was found between the DCI content in fermented flours and the biscuits prepared from them. The potential bioaccessibility of DCI from water biscuits was low. It can be concluded that, by applying select LAB and R. oligosporus 2740 for LSF, the heat treatment and physical structure of biscuits were mainly responsible for the potential bioaccessibility of DCI.

In this study, edible filamentous fungi Aspergillus oryzae and Rhizopus oligosporus were co-cultured with fodder yeast Candida utilis using corn thin stillage to produce feed-grade biomass, amylases, and ethanol with the purpose of backset valorization. The co-culture of A. oryzae with C. utilis inoculated after 24 h of pure fungal growth provided the most promising results, yielding 18.6 g L−1 biomass with 41 and 31% of lipid and protein, respectively, with all of the essential amino acids. The microorganisms consumed nearly 34% of the cultivation medium solids, as well as 33% of glycerol and 12% of lactic acid, which allowed them to produce ca. 4.5 g L−1 ethanol and 1.7 U mL−1 amylase. The liquid fraction of the cultivation medium was used as 200 mL L−1 backset for the production of high-solids corn mash. The final ethanol concentrations reached 78.3, 90.6 and 108.2 g L−1 in the control sample (without backset), unfermented backset addition and fermented backset addition, respectively. This suggests that the yeast-fungi co-culture for backset treatment was effective in improving the core production stage in an ethanol-producing plant.

The effects of solid‐state fermentation with four filamentous fungi (Rhizopus oligosporus, Actinomucor elegans, Neurospora crassa and Rhizopus oryzae) and their combination on the degradation of protein allergens and anti‐nutritional factors (ANFs) of tempeh were evaluated. Glycinin, β‐conglycinin, trypsin inhibitor and flatulence‐causing oligosaccharides were significantly hydrolysed by all assayed strains but the level of reduction of ANFs depended on the strain. ELISA was conducted to evaluate IgE immunoreactivity using sera from soy sensitive individuals. Results revealed that co‐fermentation of R. oligosporus and A. elegans resulted in the highest reduction in IgE immunoreactivity for all sera used and showed greatest degradations on α’, α, β, acidic subunits (100% hydrolysed) and extensive hydrolysis of flatulence‐causing oligosaccharides. Tempeh fermented with R. oligosporus and A. elegans displayed high level of soluble protein and peptide (

Solid-state fermentation of plant materials with food grade moulds is considered a method of obtaining bioactive food products and additives. However, the use of quinoa seeds as a substrate in such a process has not been thoroughly studied to date. In this paper, extended fermentation with Rhizopus oligosporus, Aspergillus oryzae and Neurospora intermedia (tempe, koji and oncom moulds, respectively) was performed in order to enhance the antioxidant potential of coloured quinoa (as measured by the total soluble phenols, ABTS˙+ and ˙OH scavenging activities, and the reducing power). The most efficient of the examined strains was Rhizopus, which caused an average improvement in the aforementioned qualities of 220% (black seeds) and 129% (red seeds), as compared to pre-cooked seeds. Four-day fermentation with Rhizopus resulted in a drastically enhanced amount of vanillin in the free phenolic fraction (130-fold in black quinoa seeds). However, the overall level of free and bound phenolics (as measured by the HPLC-DAD-ESI-TOF-MS method) in the material decreased. Both total peptides and individual free amino acid levels were significantly increased, which corresponded with the high antioxidant potential of the material. The fermented quinoa was also significantly enriched with total dietary fibre with dominant insoluble fraction.

During solid-state fermentation, aerial hyphae occupy the interparticle spaces, binding adjacent particles and decreasing the efficiency of mass and heat transfer within the bioreactor bed. The present work explores two questions related to this phenomenon: (i) what determines the direction of growth of vegetative aerial hyphae and (ii) what makes them stop growing? We developed a discrete mathematical model to explore the factors affecting the distribution of aerial hyphae with height above the surface and adjusted the model parameters by fitting the model to literature data of Rhizopus oligosporus growing on potato dextrose agar. The adjusted rules were simple and indicate that vegetative aerial hyphae grow in any direction, as though their tips moved in a random-walk fashion. Vegetative aerial hyphae stop extending when they enter crowded spaces, resulting in a maximum local biomass concentration. With these rules, the layer model was used to study the influence of increasing inoculum concentration and to simulate, for the first time, growth between the surfaces of two adjacent particles.

The influence of liquid-state fermentation (LSF) by selected lactic acid bacteria (LAB) and Rhizopus oligosporus fungi on the content of rutin and total phenolic compounds (TPC), antioxidant capacity measured by ABTS test, FRAP assay and photochemiluminescence technique, and the inhibitory activity against formation of fluorescent advanced glycation end-products (AGEs) in vitro of raw and roasted buckwheat flours was studied. LSF caused a slight, specific LAB-dependent increase in TPC and a decrease in rutin content. Fermented raw buckwheat flours contained higher amounts of rutin and TPC with one exception when the highest increase in TPC was noted in roasted flour fermented by fungi. A LAB-dependent difference in the antioxidant capacity of buckwheat flours was noted while the inhibitory activity of fermented flours against AGEs formation was generally reduced. It can be concluded that LSF with selected LAB and fungi may improve the antioxidant and functional properties of buckwheat flours.

The fermentation of fruit processing residuals (FPRs) with filamentous fungi can provide protein-rich food products. However, FPRs that contain bioactive compounds with antimicrobial properties present a major challenge. In this work, the resistance of two edible filamentous fungi, Rhizopus oligosporus and Neurospora intermedia, to 10 typically inhibiting bioactive compounds available in FPRs (epicatechin, quercetin, ellagic acid, betanin, octanol, hexanal, D-limonene, myrcene, car-3-ene, and ascorbic acid) was examined. These compounds’ inhibitory and stimulatory effects on fungal growth were examined individually. Three different concentrations (2.4, 24, and 240 mg/L) within the natural concentration range of these compounds in FPRs were tested. These bioactive compounds stimulated the growth yield and glucose consumption rate of R. oligosporus, while there was no increase in the biomass yield of N. intermedia. Ellagic acid caused an up to four-fold increase in the biomass yield of R. oligosporus. In addition, octanol and D-limonene showed antifungal effects against N. intermedia. These results may be helpful in the development of fungus-based novel fermented foods.

Clove essential oil is processed product of clove leaves which is used as perfumes and medicines. In general, the method used to extract clove leaf essential oil is the steam distillation method. This method has not maximized the yield essential oils produced because a lot of essential oils are still trapped in plant tissues. Modification of the essential oil isolation methods to produce more yields needs to be done, one of which is the fermentation method with the help Rhizopus oligosporus before the distillation process is carried out. The fermentation method is able to destroy the oil tissues on the leaves using enzymes produced by rhizopus oligosporus. The aims of this study were to isolate the essential oil clove leaves and look for the effect of the mass and fermentation time on the yield and content of clove leaf essential oil compounds produced. The results showed that the isolation essential oils using fermentation methods was successfully carried out, the mass and fermentation time had a significant influence on the yield and content of essential oil chemical compounds. The best result was obtained at a mass of 35 grams with a fermentation time of 36 hours with a yield 0.32% and the content eugenol, caryopillene compounds obtained respectively at 93.43% and 2.28%.

High temperature treatment and fermentation are two major preparation methods applied by consumers to make soybean more edible. Germination is a simple way to induce certain bioactive compounds in soybeans. Objective: This study aimed to determine the impact of germination on macronutrient compositions and isoflavones’ level in boiled tempe. Method: Grobogan soybean samples were sprouted at room temperature before the fermentation. The samples were fermented using 0.2% and 0.4% inoculum contained Rhizopus oligosporus before the boiling process. Fat, protein, and carbohydrate compositions were estimated by the soxhlet extraction, the Kjeldahl method, and spectrophotometric respectively. Genistein and daidzein contents in the samples were analysed by high-performance liquid chromatography (HPLC). Result: In comparison with ungerminated Grobogan tempe samples, based on dry weight, the germinated Grobogan tempe samples described a significant reduction in protein (24-hour germination period, p=0.002) and carbohydrate (24-hour and 48-hour germination periods, p

The success of formulated foodstuffs and drinks depends on the type of additives used to boost their character such as nourishment, fragrance and appearance. Some of these compounds are designated as “generally recognized as safe” (GRAS). Any constituent which is added to food and had gone through checking by the scientific professional for its intended use is regarded as GRAS. Some GRAS fungal strains have been analysed for the production of nutraceuticals. Fungal strains such as Aspergillus niger, Agaricus bisporus, Tolypocladium niveum and Rhizopus oligosporus are regarded GRAS by the Food and Drug Administration (FDA) and are used commercially in the food sector. In recent years, the knowledge of GRAS strains and their utilization in food has increased due to the easy availability of cutting-edge technologies at hand. The additional features associated with certain GRAS compounds such as odourless, neutral and flavourless can withstand a large range of temperature and pH, highlighting their positive aspects for usage in food production. Some studies have also shown that few fungal strains which are regarded as GRAS are a source of amino acids and peptides which are usually used as supplements in our diet. It is also found that some of these fungi perform duplex function: one being a healthful food ingredient and the other as flavouring agents. Hence, this chapter attempts to demonstrate why the fungal strains which are given GRAS status should be used as functional food ingredients.

Rizal S, Murhadi, Kustyawati ME, Hasanudin U. 2020. Growth optimization of Saccharomyces cerevisiae and Rhizopus oligosporus during fermentation to produce tempeh with high β-glucan content. Biodiversitas 21: 2667-2673. Saccharomyces cerevisiae grows and produces β-glucan during fermentation in tempeh production. The content of β-glucan in tempeh is influenced by the growth of S. cerevisiae throughout fermentation. The purpose of this study was to determine the effects of different types and concentrations of carbon sources on yeast growth, fungi growth, and β-glucan content in tempeh inoculated using Rhizopus oligosporus and S. cerevisiae. This study used a Factorial Randomized Complete Block Design (RCBD) with two factors and three replications. The first factor was the types of carbon sources, tapioca and wheat flour; the second factor was the concentrations of carbon source, 0.0%, 2.5%, 5.0%, 7.5% and 10.0% (w/w). Tempeh produced was investigated for yeast number, fungi number, β-glucan content, and pH value. The obtained data were tested using Tukey's Honestly Significance Difference (HSD) test. The results showed that the addition of various types and concentrations of carbon source significantly influenced the increase in yeast number, fungi number, β-glucan content, and pH in tempeh. The growth of yeast, fungi, and β-glucan content increased along with the increment of carbon source concentration. The amounts of yeast, fungi, and β-glucans in tempeh added with tapioca were higher compared to tempeh with wheat flour. The addition of 10% tapioca produced the highest amount of yeast with 9.505 Log CFU/g and the highest β-glucan content with 0.707% (w/w).

Lupin is a nutritive grain, but its use is limited due to its high content of bitter alkaloids and other antinutritional factors, such as phytic acid, tannins, nitrates and trypsin inhibitors (TI), that have undesirable physiological effects. There is increasing interest in finding appropriate methods for reducing the antinutritional compounds in lupin. The objective of this research was to assess the efficacy of a biotechnological process, namely, fungal fermentation, as a debittering process relative to that of conventional aqueous thermal treatment (ATT). We evaluated the effects of these processes on the reduction of antinutritional compounds as well as their potential impacts on enhancing the beneficial antioxidant properties of lupin. Three varieties (INIAP-450, INIAP-451 and Criollo) of the Lupinus mutabilis species were studied. The application of ATT and fermentation with Rhizopus oligosporus caused decreases in the following antinutrients: nitrates (94.59%), tannins (82.10%), alkaloids (94%), urease activity (93.75%), phytic acid (70.06%) and trypsin inhibitors (76.76%). Ascorbic acid also decreased (79.72%). All values corresponded to the average in the three varieties evaluated. While the contents of phenols, carotenoids and the antioxidant capacity decreased by 96.83, 49.42 and 96.13%, respectively, due to the debittering process, solid fermentation promoted increases in these compounds and properties in the debittered grain., This research was financially supported by the Inter-American Development Bank, (BID), Cooperative Program for the Regional Agricultural Technology Fund (FONTAGRO) (Project FTG/RF-14893-RG, 2016–2018), the Spanish Ministry of Science, Innovation and Universities (RTI2018-095919-B-C21), the European Regional Development Fund (FEDER) and the Generalitat Valenciana (Prometeo 2017/189)

Tempeh is a traditional lndonesian food produced by fermented soybeans by utilizing the fungi Rhizopus oligosporus. Tempeh is generally made from soy but can also be made pulses, which is jack bean (Canavalia ensiformis). Jack bean tempeh has a short shelf life and will immediately rot during storage. Processing of tempeh into tempeh flour is an effort to extend the shelf life of tempeh. Drying is a very important process in making flour. This would determine the quality of flour produced, because it would affect the shelf life. This study aimed to evaluate the physical, chemical, sensory characteristic and best temperature to produce jack bean tempeh flour. The tempeh was dried for 6 hours at various temperature namely 55°C, 65°C, and 75°C and then milled into flour. The results showed that the increasing of drying temperature decreasing moisture content, fat content, yield, bulk density, water absorption capacity, brightness (L*), whiteness degree, panelist’s perception of brightness, panelist’s perception of beany odorless, panelist’s acceptance for color, panelist’s acceptance on the aroma and panelist’s acceptance for the overall jack bean tempeh flour. The increasing of drying temperature increasing oil absorption capacity, redness (a*) and yellowish (b*) of jack bean tempeh flour. The treatment that produces jack bean tempeh flour with the best chemical, physical and sensory characteristic is drying with temperature of 55°C.

Bioethanol seems very potential to be an alternative energy substituting the fossil-based energy due to its renewable sources. Any sources containing carbohydrate (poly or mono) may be fermented into ethanol (bioethanol). This study aims to investigate the optimum conditions in the fermentation of used paper HVS hydrolysate to yield ethanol. The used paper HVS hydrolysate, produced from the hydrolysis of used paper HVS, was initially treated with a so-called detoxification stage to remove compounds hindering the fermentation process. The detoxification stage employed adsorptive separation with the aid of carbon (charcoal) activated with KOH and K2CO3 solution. The fermentation varied the types of microbes or yeast (i.e. Saccharomyces cerevisiae, Rhizopus oligosporus) and its concentration. Within the ranges of studied parameters, the optimum conditions to yield highest level of ethanol (0.436%) were activated carbon with KOH, yeast of Saccharomyces cerevisiae, and concentration of 3 g yeast/70 mL hydrolysate.

The aims of this study were to determine the physico-chemical features of tempehs produced from different sources of legume (soybean (control), chickpea, lentils (red and green), and beans (white, black, and broad)) using Rhizopus oligosporus and to specify the best tempeh by sensory evaluation. Results showed that the lowest ash (0.53%) and moisture (44.88%), and the highest protein (47.11 mg/mL) levels were determined in chickpea tempeh, which contained slightly low phenolic compared to soybean tempeh while its sugar content was about 3.58-fold higher than that of soybean tempeh. Thus, the moisture content of white bean (55.20%) and red lentil (55.99%) tempehs was similar to soy tempeh (55.45%), and the ash contents of the remaining tempehs except for chickpea tempeh (0.53%) were approximately the same (1.20–1.91%). The color and texture profile values of chickpea tempeh showed similarity with that of soybean tempeh. Besides, while no phytic, acetic, fumaric, and iso-citric acids were detected in chickpea, red lentil and broad bean tempehs, acetic acid was detected in soybean, white bean, black bean, green lentil, and broad bean tempehs. Sensory evaluations indicated that the chickpea tempeh was superior to the other produced tempehs. Consequently, tempehs from different sources of legume were successfully produced.

Today the main source of unsaturated fatty acids especially omega-3 is still from marine fish oil. In this research, Rhizopus oligosporus will be used to produce PUFA (polyunsaturated fatty acids). This research used submerged fermentation method and temperature shift strategy to optimize the production of PUFA. In this result, batch culture of R.oligosporus for PUFA production at various temperatures (22, 26, 32, and 37°C). Result showed that R.oligosporus has different optimum temperature for growth and PUFA production, 26°C and 22°C respectively. Based on the result temperature-shift strategy was applied, in which the culture temperature was controlled at 26°C at the first 5 days, and then switched to 22°C for two days. By applying such a temperature-shift strategy, the dry cell weight decreased from 0,55 g to 0,47 g and the concentration of PUFA increased from 19,8% to 22,5%.

Cardiovascular disease is one of the biggest killing diseases in the world especially in Indonesia. The percentage risk of people stricken by this disease is so high in community and even causing death. This risk can actually be prevented with dietary addition of consuming unsaturated fatty acid that id often found in fish oil. However, the availability and quality of the fish in Indonesia's ocean is such that nowadays pollution has become the biggest threat. With the problem above, the alternative sources of omega 3 and omega 6 are needed whereas the source must be able to produce lipids in large number without taking up a large amount of land and also economic resources. There are several kinds of microorganisms that can be used as an alternative source of fatty acid. In this research, Rhizopus oligosporus is used as fungi microorganism. Mold is used in this research, because it has a lot of advantages such as ease to handle, easily grown in low pH and able to degradate more complex carbon sources. In this research, the fermentation is processed with submerged fermentation with temperature shift strategy with variation of glucose concentration and agitation rate. The results showed that the unsaturated fatty acid content obtained in the optimum conditions both from the glucose concentration of 100 g / L and agitation rate of 120 rpm are 41.77% in MUFA and 13.41% in PUFA and the highest highest oleic and linoleic acids were also obtained at 41.06% and 13.41%.