Optimization of Protease and Amylase Production by Rhizopus oryzae Cultivated on Bread Waste Using Solid-State Fermentation

This research was carried for the coproduction of two industrial enzymes: α-amylase and protease via SSF by Rhizopus oryzae on humidified bread waste. Fermentation time, inoculum size, initial moisture content, salt solutions, and the thickness of the substrate were investigated one by one. Fungus culture was carried out in sterile aluminum trays, and pH was adjusted to 5.5. The main results showed that the highest levels of enzyme production were obtained at 120 h, 65% relative humidity, height media of 1 cm, 105 spore/g, and M-9 solution (g/L): NaH2PO4, 12.8; KH2PO4, 3; NaCl, 0.5; NH4Cl, 1; MgSO4 7H2O, 0.5; CaCl2 2H2O, 0.01. α-Amylase (100 U/g) and protease (2400 U/g) produced by SSF from Rhizopus oryzae (CH4) on BW as substrate are of great interest in industries and could be valorized as enhancers of the bread making process.

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Production of thermostable extracellular α-amylase by a moderate thermophilic Bacillus licheniformis isolated from Qinarje Hot Spring (Ardebil prov. of Iran)

Periodicum Biologorum ◽

10.18054/pb.v118i4.3737 ◽

2017 ◽

Vol 118 (4) ◽

Cited By ~ 4

Author(s):

Ali Deljou ◽

Iman Arezi

Keyword(s):

Carbon Source ◽

Nitrogen Source ◽

Bacillus Licheniformis ◽

Bacterial Growth ◽

Enzyme Production ◽

Hot Spring ◽

Basal Medium ◽

Thermophilic Bacterium ◽

Inoculum Size ◽

Amylase Production

Background and Purpose: Amylases are most important industrial enzymes that account for about 30% of the world’s food, feed, fermentation, textile, detergent and cellulosic industries. This study aimed at optimum production of thermostable α-amylase via moderate thermophilic bacterium (Bacillus licheniformis) which was recently isolated from Qinarje Hot spring.Material and Methods: Initially, ability of bacterium for amylase activity was determined by starch hydrolysis test using Gram’s iodine staining. Then bacterial growth pattern and amylase production curves in basal production medium were graphically determined at different time intervals. Finally, effect of different temperature, pH, carbon source, nitrogen source, minerals and inoculum size were studied on bacterial growth and amylase production using turbidimetric and DNS method, respectively.Results: Optimum enzyme production achieved after 84 hours of inoculation from cultures growing at 40 ˚C and pH 9.0 in a medium containing 0.03% (w/v) of CaCl2, compared to the basal medium, results showed that the best enzyme production happened with inoculum size of 4% (v/v). The addition of 1% (w/v) rice husk (as a Carbon source) enhanced enzyme productivity up to 160% and substitution of the peptone and yeast extract with 1% (w/v) of tryptone (as a Nitrogen source) increased the α-amylase production up to 160%.Conclusion: Our findings show that B. licheniformis-AZ2 strain has an ability to produce the thermostable α-amylase which is suitable in starch processing and food industries. To be commercialized, further investigation is required for enhancement of the enzyme production.Keywords: Bacillus licheniformis; Optimization; Basal medium; Agricultural by-products.

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β-Galactosidase-Producing Isolates in Mucoromycota: Screening, Enzyme Production, and Applications for Functional Oligosaccharide Synthesis

Journal of Fungi ◽

10.3390/jof7030229 ◽

2021 ◽

Vol 7 (3) ◽

pp. 229

Author(s):

Bettina Volford ◽

Mónika Varga ◽

András Szekeres ◽

Alexandra Kotogán ◽

Gábor Nagy ◽

...

Keyword(s):

Wheat Bran ◽

Enzyme Production ◽

Specific Activity ◽

Prebiotic Synthesis ◽

Crude Enzyme ◽

Oligosaccharide Synthesis ◽

Industrial Enzymes ◽

Chromogenic Agar ◽

Lichtheimia Ramosa ◽

Excellent Source

β-Galactosidases of Mucoromycota are rarely studied, although this group of filamentous fungi is an excellent source of many industrial enzymes. In this study, 99 isolates from the genera Lichtheimia, Mortierella, Mucor, Rhizomucor, Rhizopus and Umbelopsis, were screened for their β-galactosidase activity using a chromogenic agar approach. Ten isolates from the best producers were selected, and the activity was further investigated in submerged (SmF) and solid-state (SSF) fermentation systems containing lactose and/or wheat bran substrates as enzyme production inducers. Wheat bran proved to be efficient for the enzyme production under both SmF and SSF conditions, giving maximum specific activity yields from 32 to 12,064 U/mg protein and from 783 to 22,720 U/mg protein, respectively. Oligosaccharide synthesis tests revealed the suitability of crude β-galactosidases from Lichtheimia ramosa Szeged Microbiological Collection (SZMC) 11360 and Rhizomucor pusillus SZMC 11025 to catalyze transgalactosylation reactions. In addition, the crude enzyme extracts had transfructosylation activity, resulting in the formation of fructo-oligosaccharide molecules in a sucrose-containing environment. The maximal oligosaccharide concentration varied between 0.0158 and 2.236 g/L depending on the crude enzyme and the initial material. Some oligosaccharide-enriched mixtures supported the growth of probiotics, indicating the potential of the studied enzyme extracts in future prebiotic synthesis processes.

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Lovastatin Production byAspergillus terreusUsing Agro-Biomass as Substrate in Solid State Fermentation

Journal of Biomedicine and Biotechnology ◽

10.1155/2012/196264 ◽

2012 ◽

Vol 2012 ◽

pp. 1-11 ◽

Cited By ~ 27

Author(s):

Mohammad Faseleh Jahromi ◽

Juan Boo Liang ◽

Yin Wan Ho ◽

Rosfarizan Mohamad ◽

Yong Meng Goh ◽

...

Keyword(s):

Particle Size ◽

Solid State ◽

Moisture Content ◽

Nitrogen Source ◽

Solid State Fermentation ◽

Incubation Temperature ◽

Initial Moisture Content ◽

Fermentation Time ◽

Maximum Production ◽

Additional Nitrogen

Ability of two strains ofAspergillus terreus(ATCC 74135 and ATCC 20542) for production of lovastatin in solid state fermentation (SSF) using rice straw (RS) and oil palm frond (OPF) was investigated. Results showed that RS is a better substrate for production of lovastatin in SSF. Maximum production of lovastatin has been obtained usingA. terreusATCC 74135 and RS as substrate without additional nitrogen source (157.07 mg/kg dry matter (DM)). Although additional nitrogen source has no benefit effect on enhancing the lovastatin production using RS substrate, it improved the lovastatin production using OPF with maximum production of 70.17 and 63.76 mg/kg DM forA. terreusATCC 20542 andA. terreusATCC 74135, respectively (soybean meal as nitrogen source). Incubation temperature, moisture content, and particle size had shown significant effect on lovastatin production (P<0.01) and inoculums size and pH had no significant effect on lovastatin production (P>0.05). Results also have shown that pH 6, 25°C incubation temperature, 1.4 to 2 mm particle size, 50% initial moisture content, and 8 days fermentation time are the best conditions for lovastatin production in SSF. Maximum production of lovastatin using optimized condition was 175.85 and 260.85 mg/kg DM forA. terreusATCC 20542 and ATCC 74135, respectively, using RS as substrate.

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Biochemistry of postharvest spoilage of sweet potato (Ipomoea batatas L.). 2. Comparison of cellulolytic enzyme production in cultures and fungi-infected sweet potato tubers

Annals of Tropical Research ◽

10.32945/atr2814.2006 ◽

2006 ◽

pp. 48-57

Author(s):

R. C. Ray

Keyword(s):

Sweet Potato ◽

Enzyme Production ◽

Ipomoea Batatas ◽

Rhizopus Oryzae ◽

Cellulose Synthesis ◽

Cellulolytic Enzyme ◽

Optimum Temperature ◽

Potato Tubers

The study was conducted to determine the production in vitro and in vivo of cellulases by Botrydiplodia theobromae and Rhizopus oryzae. Isolates of these organisms were obtained from the postharvest decay of sweetpotato tubers. Results revealed that B. theobrornae and R. oryzae which were isolated from postharvest spoilage of sweetpotato tubers produced endo-13-1,4-glucanase and exo-V-1 ,4-glucanase in culture and in fungi-infected tissues of sweetpotato tubers. The optimum temperature and pH for cellulose synthesis and activity were 30°C and pH 6.5, respectively.

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Optimal Conditions For Production Acidic D-Xylanase By Aspergillus niger in Submerged Fermentation

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2011.5.3.174 ◽

2011 ◽

Vol 5 (3) ◽

pp. 14-21

Author(s):

Muhamed Omar Abdulatif ◽

Hyder H. Assmaeel ◽

Raghad kadhim Obeid ◽

Ayat Adnan Abbas

Keyword(s):

Aspergillus Niger ◽

Rice Husk ◽

Enzyme Production ◽

Inoculum Size ◽

Optimal Conditions ◽

Optimum Conditions ◽

Xylanase Production ◽

Primary Isolation ◽

Optimum Ph ◽

Optimum Period

he Xylanase producing strain Aspergillus niger was isolated from soil on potato dextrose agar in the presence of xylan as its first substrate for primary isolation, and then grown under liquid medium fermentation in the presence of crude xylan (rice husk) to produce D-Xylanase. the optimum conditions were determined as follows: the Optimum pH for xylanase production was found pH 5.0, xylanase was induced by xylan (rice husk) 0.1% and the production was (61.221 U/ml) and nitrogen source Yeast extract recorded highest enzyme production( 89.71 U/ml), and repressed by carbon source xylose the highest enzyme production (88.69 U/ml). The optimum temperature was 40°с for xylanase production was (35.15 U/ml), the optimum period after 7 days of incubation was (52.33 U/ml) ,the optimum substrate concentration 0.1% was (45.95 U/ml), and the optimum inoculum size was 1 x 106 (spore /ml) recorded (57.19 U/ml ).

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Improvement of the Quality of Ginkgo biloba Leaves Fermented by Eurotium cristatum as High Value-Added Feed

Processes ◽

10.3390/pr7090627 ◽

2019 ◽

Vol 7 (9) ◽

pp. 627

Author(s):

Minmin Zou ◽

Xiaohan Guo ◽

Yan Huang ◽

Fuliang Cao ◽

Erzheng Su ◽

...

Keyword(s):

Ginkgo Biloba ◽

Value Added ◽

Inoculum Size ◽

Bioactive Substances ◽

Fermentation Time ◽

Nutrient Changes ◽

Unpleasant Smell ◽

Optimal Medium ◽

Ginkgolic Acids

Ginkgo biloba leaves are well known for their high content of nutrients and bioactive substances. However, unpleasant smell and a small number of ginkgolic acids greatly reduce the utilization of the leaves. In this work, solid-state fermentation of G. biloba leaves using Eurotium cristatum was studied by investigation of the nutrient changes and its feasibility as a functional feed. E. cristatum could grow on pure G. biloba leaves and the addition of excipients could significantly improve the growth of E. cristatum. The optimal medium was with 10% (w/w) of whole G. biloba seeds and the optimized water content, pH, inoculum size and fermentation time were 45% (w/w), 4.5, 4.76 × 107 CFU/100 g wet medium, and eight days, respectively. Under the optimal conditions, the spore number increased by about 40 times. The content of flavonoids was greatly increased by 118.6%, and the protein and polyprenyl acetates (PPAs) were increased by 64.9% and 10.6%, respectively. The ginkgolic acids, lignin, and cellulose were decreased by 52.4%, 38.5%, and 20.1% than before, respectively. Furthermore, the fermented G. biloba leaves showed higher antioxidant activity and held more aroma substances. Thus, G. biloba leaves fermented by E. cristatum have potential as s high value-added feed. This is the first investigation of E. cristatum fermentation on ginkgo leaves, which will facilitate the use of ginkgo leaves in the feed industry.

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Production and application of amylases of Rhizopus oryzae and Rhizopus microsporus var. oligosporus from industrial waste in acquisition of glucose

Chemical Papers ◽

10.2478/s11696-013-0466-x ◽

2014 ◽

Vol 68 (4) ◽

Cited By ~ 13

Author(s):

Aline Freitas ◽

Bruna Escaramboni ◽

Ana Carvalho ◽

Valéria Lima ◽

Pedro Oliva-Neto

Keyword(s):

Rhizopus Oryzae ◽

Industrial Wastes ◽

Fermentation Condition ◽

Rhizopus Microsporus ◽

Batch Cultures ◽

Cassava Bagasse ◽

Glucose Syrup ◽

Amylase Production ◽

Ph Range ◽

Better Than

AbstractAmylases from Rhizopus oryzae and Rhizopus microsporus var. oligosporus were obtained using agro-industrial wastes as substrates in submerged batch cultures. The enzymatic complex was partially characterised for use in the production of glucose syrup. Type II wheat flour proved better than cassava bagasse as sole carbon source for amylase production. The optimum fermentation condition for both microorganisms was 96 hours at 30°C and the amylase thus produced was used for starch hydrolysis. The product of the enzymatic hydrolysis indicated that the enzyme obtained was glucoamylase, only glucose as final product was attained for both microorganisms. R. oligosporus was of greater interest than R. oryzae for amylase production, taking into account enzyme activity, cultivation time, thermal stability and pH range. Glucose syrup was produced using concentrated enzyme and 100 g L−1 starch in a 4 hours reaction at 50°C. The bioprocess studied can contribute to fungus glucoamylase production and application.

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Isolation of Thermostable Protease Producing Bacteria from the Microbial Mats

Journal of University of Shanghai for Science and Technology ◽

10.51201/jusst/21/09703 ◽

2021 ◽

Vol 23 (10) ◽

pp. 222-235

Author(s):

Malathi, M ◽

◽

Latha, D ◽

Keyword(s):

Enzyme Production ◽

Microbial Mats ◽

Skim Milk ◽

Inoculum Size ◽

Protease Production ◽

Carbon And Nitrogen ◽

Test Culture ◽

Higher Temperature ◽

Optimized Conditions ◽

Near Future

As thermostable protease has more commercial value in different industries, the aim of this study was to search for such an enzyme producing bacteria from the microbial mats. Investigation was continued on the isolate for its ability to produce mass amount of enzyme and its activity under suitable optimized conditions. Different parameters including cheap carbon and nitrogen substrates, inoculum size and temperature was selected to optimize the enzyme production conditions. Initially five different isolates from two microbial mats collected from different sources were analyzed for its ability to produce thermostable protease after exposing to higher temperature incubation conditions. Test culture tentatively named as 1F from microbial mat-1 was selected as more enzyme producer among the ten isolates. The organism was selected based on the zone of clearance on skim milk agar by the isolate, that indicating more protease production. Under each optimization parameter, each type of carbon (Lactose), and nitrogen (yeast extract) source showed more enzyme production and activity respectively. About 1% inoculum size and a thermostable temperature of 45°C produced significant amount of enzyme and its activity. The obtained results emphasized the need for thermostable protease for different commercial industries in the existing and near future.

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Milk-clotting proteases from Pleurotus albidus: an innovative alternative for the production of Minas frescal cheese

Acta Scientiarum Biological Sciences ◽

10.4025/actascibiolsci.v43i1.57275 ◽

2021 ◽

Vol 43 ◽

pp. e57275

Author(s):

Salomão Rocha Martim ◽

Larissa Svetlana Cavalcante Silva ◽

Mircella Marialva Alecrim ◽

Lorisa Simas Teixeira ◽

Maria Francisca Simas Teixeira

Keyword(s):

Temperature Stability ◽

Iodoacetic Acid ◽

Inoculum Size ◽

Cultivation Medium ◽

Fermentation Time ◽

Milk Clotting ◽

Inoculum Age ◽

Initial Ph ◽

Promising Source ◽

Stability Effect

Pleurotus albidus, a naturally growing species in the Amazon region, has been considered a promising source of milk-clotting proteases. The production of such enzymes using lignocellulosic residues is a sustainable alternative to replace mammalian rennet. The application of P. albidus milk-clotting proteases in cheese making has not yet been reported in the scientific literature. The aim of this study was to characterize the milk-clotting proteases of P. albidus and use these enzymes in the production of Minas frescal cheese. For the production of coagulating proteases, the mushroom was grown in açaí seeds supplemented with rice bran (10%, w/w). The parameters affecting the production of coagulant, such as inoculum size, fermentation time, initial pH of cultivation medium and age of the inoculum were evaluated. The coagulant extract obtained under optimal production conditions was evaluated for optimal pH and temperature, pH and temperature stability, effect of ions and inhibitors. Significant production of coagulating proteases was obtained under the following conditions: inoculum size (2.5%), fermentation time (10 days), initial pH of the cultivation medium (6), and inoculum age (10 days). The coagulant exhibited significant catalytic activity in pH 5.0 at 55°C, with stability at 45°C and was completely inhibited by iodoacetic acid. The milk-clotting proteases of P. albidus were efficient for making Minas frescal cheese that presented 55.0% of moisture, 20.0% of lipids and 17.20% of protein. Pleurotus albidus is a potential source of milk-clotting proteases that can be applied in dairy industry for production of fresh Minas frescal cheese.

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OPTIMIZATION OF CELLULASE ENZYME PRODUCTION FROM Aspergillus oryzae FOR INDUSTRIAL APPLICATIONS

World Journal of Biology and Biotechnology ◽

10.33865/wjb.002.02.0088 ◽

2017 ◽

Vol 2 (2) ◽

pp. 155 ◽

Cited By ~ 1

Author(s):

Hassan Sher ◽

Muhammad Faheem ◽

Abdul Ghani ◽

Rashid Mehmood ◽

Hamza Rehman ◽

...

Keyword(s):

Enzyme Production ◽

Time Course ◽

Value Added ◽

Cellulase Production ◽

Industrial Applications ◽

Inoculum Size ◽

Cellulosic Biomass ◽

High Yield ◽

Carboxymethyl Cellulase ◽

Cellulase Enzyme

Cellulases are the hydrolytic group of enzymes, responsible for release of sugars in the bioconversion of the cellulosic biomass into a variety of value added industrial products. Fungal isolated cellulases are well studied and playing a significant role in various industrial processes. Enzymatic depolymerisation of cellulosic material has been done by the various fungal isolated enzymes. In the present study, the cultivation conditions for cellulase production from Aspergillus species were optimized. Optimization of scarification conditions such as time course, inoculum size, carbon source and concentration, nitrogen source, various pH levels were performed for the production of extracellular carboxymethyl cellulase and endoglucanase enzyme. The result exhibited, 15 % inoculums size, corncobs 2 % concentration, Urea and medium pH 7 at 30oC supported high yield of carboxymethyl cellulase (38.80 U/ml/min) and exoglucanase enzyme (10.94 U/ml/min) through a submerged fermentation (SmF). In future biotechnological applications in cellulase enzyme production attain a vital role to obtain high degradable yield.

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