Submerged Fermentation Research Articles

Purification and characterization of novel α-amylase from Anoxybacillus ayderensis FMB1

The thermophilic amylase producing Anoxybacillus ayderensis FMB1 was isolated from Sorgun Hot Spring in Turkey. In order to obtain improved production of the enzyme for cost-effective process, optimization of the process parameters is required. Optimum conditions for amylase production were determined using the traditional Submerged Fermentation (SmF) method. The maximum amylase activity was 295.8 U/mg, under the optimized conditions (24-h, 55 °C, and pH 8.0) in liquid BMB medium. The best enzyme production was obtained in the presence of 1.5% soluble starch as carbon source and 1% yeast extract as nitrogen source. Enzyme was produced under optimal conditions and partially purified. Partially purified enzyme was stable for up to 120 minutes at 50 °C, and its stability was found to be pH 5.0–10.0. Regarding the effects of some metals and chemicals on the activity of the partially purified enzyme, Ca2+ increased the activity by 21%, but enzyme activity was decreased by Mg2+, Zn2+, Ni2+, and Cu2+ by 5%, 29%, 57%, and 64%, respectively. and the enzyme activity was completely inhibited by Hg2+. In addition; DTT, PMSF, PCMB, SDS and EDTA caused a decrease in enzyme activity by 3%, 4%, 12%, 27% and 77%, respectively. The molecular weight of enzyme was found to be 58.5 kDa by SDS-PAGE. The results obtained show that the thermophilic A. ayderensis FMB1 and their enzyme have biotechnological importance.

Effect of Type of Nitrogen Sources and Concentration on Protease Production: Fish Waste Hydrolysate as a Potential Low Cost Source

Fish waste hydrolysate is obtained from hydrolysis process of a fishery industry waste. It can be a good substrate source for the production of protease via a fermentation process. Protease covers above than 50% of the global industrial enzymes market and is regarded as an enzyme which is widely used in many applications. This research aims to determine the effect of the type of nitrogen sources and nitrogen source concentration on protease production from the fish waste hydrolysate. The experiments were done using one factor at a time (OFAT) of type of nitrogen sources and nitrogen source concentration in submerged fermentation (SmF) of 10% (v/v) Bacillus cereus (ATCC 10876). In the first stage, the variation of type of 1% (w/v) nitrogen sources such as fish waste hydrolysate, beef extracts, yeast extracts, tryptone, peptone and skim milk powder that mixed with the basal media were compared to evaluate the highest protease production. Then, the concentration of fish waste hydrolysate was varied from 1% to 3% (w/v) to analyse the production trend of protease. The protease activity and total protein content were then analysed by protease assay using tyrosine as a standard and Bradford method, respectively. Upon the observation, 1% (w/v) nitrogen sources on protease production revealed that the fish waste hydrolysates and beef extracts were capable to produce a high amount of protease (23.659 ± 0.38 U/mL and 28.7631 ± 0.59 U/mL, respectively). Among the concentration tested, 3% (w/v) of fish waste hydrolysate was found to be the highest protease production (190.85 ± 0.38 U/mL). Therefore, the fish waste hydrolysate has a potential as a low-cost source for protease production.

Optimization of Lovastatin Production from A Local Isolate of Aspergillus terreus A50 in Solid State Fermentation by Classical and Statistical Methods

Lovastatin is one of the most important compounds that is produced from some filamentous fungi, being employed in the reduction of hypocholesterolemia. The results of screening, after the collection of seventy-three local fungal isolates from different areas, demonstrated that the local isolate Aspergillus terreus A50 was the best isolate for lovastatin production, with a concentration of 12.66 µg/ml, through the submerged fermentation. Lovastatin produced from A. terreus A50 showed antimicrobial activities against a Candida albicans isolate. Solid state fermentation (SSF) was the best system to produce the highest yield of lovastatin by A. terreus A50 as compared to the submerged fermentation (SmF) system, with and without agitation. The optimum conditions for lovastatin production by SSF were also determined. The parameters included carbon sources (wastes), carbon sources mixture, incubation temperature, and moisturizing solution, which are commonly used in classical procedures. The results showed that a higher lovastatin production of 102.321 µg/gm substrate was obtained in the culture containing wheat bran and oat bran (1:1 w:w), sodium acetate, moisture ratio of 1.2 v:w, pH 7, incubation temperature of 30 °C and incubation period of 6 days. Some of these parameters, including pH, incubation period, and moisture ratio were determined by utilizing the Response Surface Method (RSM) as a statistical approach.

Isolation and Screening of Fungal Culture Isolated From Algerian Soil for the Production of Cellulase and Xylanase

Lignocellulolytic enzymes constitute a very large group of extracellular proteins secreting by fungi who is ecologically involved in the degradation of a variety of complex materials, a property that is attributed to a battery of enzymes produced by these microorganisms like cellulases and xylanases who are of significant industrial value and relevance. Forty fungal isolated from rich soil in organic matter were screened for lignocellulolytic enzymes production, its organized on the basis of their hydrolytic potential of cellulose and xylan. The isolates strains presented enzymatic activity which was ranked as follows: cellulolytic (56%), xylanolytic (44%). Some selected strains that produce high levels of enzymes (cellulase, xylanase) grown in submerged fermentation (SmF) and were quantitatively evaluated. The fermentation experiments were carried out in shake flasks. The highest CMCase (5,10 IU/ml) and xylanase (98,25 IU/ml) activities were obtained from Trichoderma sp strain Mtr6 isolate.
 Keywords: Fungi, Trichoderma sp, lignocellulolytic enzymes, soil, screening, organic matter.

Recovery of melon residues (Cucumis melo) to produce lignocellulolytic enzymes

Lignocellulosic biomass is considered the raw material of the future to produce ethanol, due to its low cost and its enormous availability. Lignocellulosic enzymes play a crucial role in the costs and sustainability of biorefining processes by promoting the biomass fractionation. In this study, the melon waste was used as inducer source to produce lignocellulolytic enzymes by Aspergillus niger M4 strain under submerged fermentation (SmF). The chemical characterization of the whole fruit showed a high content of sugars, as well as hemicellulose, followed by lignin and cellulose. The dried and crushed melon was used as a substrate in submerged fermentation, to induce the synthesis of lignocellulosic enzymes. The Aspergillus niger M4 strain was able to grow and synthesize lignocellulose degrading enzymes, allowing free-glucose values of 3.7 g/L. The culture broth showed endo-glucanase activity (1.21 U/mL), xylanase activity (11.00 U/mL), and laccase activity (18.23 U/mL). The entire melon (Cucumis melo L.) wastes can be used as a substrate in the lignocellulosic enzyme production by submerged fermentation process for further application in the biomass pretreatment for bioethanol production.

Production of Cellulase and Xylanase from Eupenicillium Javanicum by Solid-State Fermentation Utilizing Pineapple Crown Leaves Waste as the Substrate

Vast applications of cellulases and xylanases in many fields over the last few decades and high production cost demand extensive research in improving its quality and finding low cost substrates. These includes pulp and paper, food and feed, brewing, agriculture and biofuel industries. The main objective of this study was to investigate the use of pineapple crown leaves waste as the substrate to produce cellulases and xylanases in solid-state cultures (SSF) of Eupenicillium javanicum InaCC F154. Three SSF temperatures (25°C, 30°C and 35°C) and pH (4.0, 6.0 and 8.0) were tested for 96 h in solid media containing defatted soybean, (NH4)2SO4, NaNO3, CaCl2, and small amount of H2O. Then, its comparison with submerged fermentation (SmF) was also carried out. The SSF results showed that increasing temperature from 25°C to 35°C, increased the production of these enzymes. The pH also affected its activities, except at pH 4.0. The optimum pH and temperature for cellulase and xylanase production were found to be 6.0 and 35°C, respectively. The maximum cellulase obtained was 0.261 U/mL while much higher was obtained for xylanase with activity of 1.683 U/mL. SSF was better than SmF for xylanases (0.501 U/mL vs. 1.683 U/mL). The outcome of this study exhibits the potential of pineapple crown leaves as the substrate for xylanase production from E. javanicum by solid-state fermentation.

Bioconversion of lignocellulosic substrates for the production of polyhydroxyalkanoates

Lignocellulose, the most abundant and renewable carbon resource, is untapped due to its protective lignin shield. Biological pretreatment employs white rot fungi (WRF) to remove lignin and expose the cellulose/hemicellulose layers for exploitation. The present work was conducted to explore the possibility of biologically pretreating lignocellulose with WRF and using the sugars, liberated from this process, as renewable and inexpensive feedstock for the production of polyhydroxyalkanoates (PHA). Pleurotus ostreatus MTCC 142 (P142) was used to bring about bio-delignification of lignocellulose under solid state fermentation (SSF) conditions. The hydrolysates thus obtained were used for bioconversion to PHA, using Bacillus megaterium Ti3 under submerged fermentation (SmF) conditions in a nitrogen-deficient medium. From the lignocellulosic substrates used, the highest sugar yields, post bio-delignification, were obtained from corn husk (6 mg/mL). Production of PHA from pure sugars revealed a preference for xylose with 50% PHA accumulation. The pretreated corn husk (CH) hydrolysate yielded 1 g/L PHB with an accumulation of 57.8% after 48 h, comparable to that obtained using xylose based medium and significantly higher (p < 0.05) than the PHA accumulation (45.7%) obtained from the glucose medium. Fermentation kinetics revealed that PHB yield obtained from the hydrolysate was 3 fold higher than glucose-based production. Similarly, PHB yield, in terms of substrate conversion Yp/s (gPHA/gS), also increased 2 folds in the case of biologically pretreated lignocellulosic bioconversion to PHB. Characterization of the polymer through Proton (1H) and Carbon (13C) NMR analyses revealed that it was the short-chain-length (scl) polyester, poly-β-hydroxybutyrate (PHB).

Plant-based proteinaceous snacks: Effect of fermentation and ultrasonication on end-product characteristics.

The study aimed at the development of a sufficient technology to improve sensory, textural, physical, and microbiological properties of peas snacks (Ps) using solid‐state fermentation (SSF) and submerged fermentation (SMF) with two different lactic acid bacteria (LAB) strains (Lactobacillus casei LUHS210 and Lactobacillus uvarum LUHS245) for 24 hr and ultrasonication (10, 20, and 30 min). To ensure safety of the used technologies, microbiological characteristics and biogenic amines (BAs) content in treated Ps were analyzed. Additionally, a different salt content (3.6 and 1.0 g/100 g) was used for snacks preparation. The obtained results revealed that used treatments reduced enterobacteria in Ps, while in fermented Ps, yeast/moulds were not found. Ps with the lower salt content were more acidic and harder (0.90 mJ), and there was a significant effect (p < .05) due to the fermentation method, LAB strains, and ultrasonication on the texture of final product. Different salt content significantly affected the color coordinates of the Ps tested (p < .05). The predominant biogenic amines in Ps were phenylethylamine and spermidine. However, the reduction of some BAs after samples fermentation was observed. To conclude, acceptable formulations of Ps can be obtained with 1.0 g/100 g salt, and by using fermentation, as the end‐product is more attractive to consumers than those prepared with 3.6 g/100 g salt and using ultrasonication.

Statistical Optimization of Xylanase from Bacillus licheniformis Using Banana Peels in Submerged Fermentation

Five previously isolated bacterial strains were cultured on xylan agar medium (XAM) to screen their xylanase production potential. Two best strains were obtained on the basis of clear zones on XAM. The strain, Bacillus licheniformis, produced higher xylanase (3.24 U/ml) was selected for further studies based on quantitative xylanase assay. Both cultural and medium parameters were optimized to yield highest xylanase production. Best cultural conditions found were 5 pH, 1% inoculum size, 37 °C and 24 h of fermentation period. To optimize the xylanase production, medium ingredients viz, banana peels, yeast extract, KH2PO4, K2HPO4, sodium nitrate, MgSO4, peptone, NaCl and ammonium sulfate were screened through 12 runs Plackett–Burman design of response surface methodology. From 12 experimental runs, 4 significantly affecting medium ingredients (banana peels, sodium nitrate, peptone, MgSO4) were selected and further optimized through central composite design. The banana peels (1.5%), sodium nitrate (0.3875%), peptone (0.8%) and MgSO4 (0.775%) had yielded significantly highest xylanase production. Partial characterization of crude xylanase revealed optimum 8 pH and 55 °C temperature.

Amylase Production by Bacillus subtilis SY134D Strain Under Submerged Fermentation

Industrial important enzymes have traditionally been obtained by using submerged fermentation technology. Many organisms are able to produce these enzymes, but only a few of them exhibit satisfactory characteristics for industrial application. The ability of bacterial strains from the genus Bacillus to secrete large amounts of extracellular a-amylase has made them well suited for commercial production. Amylase production using Bacillus subtilis SY134D strain was conducted in flasks and bioreactor. In flasks, our results showed that soybean cake, beet pomace and tomato pomace were the best carbon source used in submerged fermentation (SmF) after 72 h of incubation. In bioreactor, the rates of air flow and agitation speed are important factors that affect oxygen transfer rate which influence on product formation. When the aeration rate was increased from 0.25 to 0.75 vvm, amylase production increased at the speed of agitation 100 and 200 rpm. The best enzyme yield 127 IU/mL was obtained at 0.25 vvm an aeration rate and 300 rpm agitation speed in a 3L Electro-lab bio-reactor.&#x0D; &#x0D; &#x0D;

Safety assays and nutritional values of mycoprotein produced by Fusarium venenatum IR372C from date waste as substrate.

Nutritional and environmental benefits of mycoprotein verify its beneficial role on the health of humankind in the next decades. Agro-industrial wastes can be used as cheap substrates to decrease the total cost of product. However, fungi may produce toxins or lead to allergic reactions in consumers. Therefore, the study of the safety and nutritional aspects of this product are very important. Fusarium venenatum IR372C was cultured on date wastes and ammonium salts in submerge fermentation. The safety and nutritional issues of produced mycoprotein were investigated including allergy tests and analyses of toxins, as well as existence of toxin genes, and content of heavy metals, metals, amino acids and fatty acids. The results showed that fumonisin genes in F. venenatum IR372C remain without any gene expression during 1 week fermentation. Zearalenone and deoxynivalenol cannot be detected in the fermentation medium after 3 weeks. Prick tests on 30 volunteers demonstrated no sensitivities to mycoprotein. The content of lead was 658 μg kg-1 as the highest heavy metal followed by arsenic, cadmium and mercury at 161, 30.57 and 0 μg kg-1 , respectively. Produced mycoprotein includes essential amino acids at appropriate contents and the ratio of unsaturated to saturated fatty acid was nearly 2:1. Also, calcium, iron, magnesium and zinc were found in mycoprotein, which could have health beneficial impacts on consumers. This study has provided information on safety aspects of mycoprotein production by F. venentaum IR372C from date wastes. However, further studies with focus on long-term clinical benefits of diets containing mycoprotein are necessary. © 2020 Society of Chemical Industry.

A critical review on submerged production of mushroom and their bioactive metabolites.

Mushrooms are ubiquitous in nature. Even though humankind has been consuming mushrooms for ages, their medicinal and nutraceutical properties are not used to its fullest potential in the present market. Edible mushrooms are not only a cheap and nutritious option to mitigate malnutrition, but they also produce effective biomass. Submerged fermentation (SmF) is not only a cost-effective method to produce biomass along with exquisite bioactive metabolites but it also reduces the chances of contamination and the time of production. Therefore, this study unveils the bioactive metabolites being produced by mushrooms. Moreover, it also showcases the recent advances in the areas of bio-active compounds and their judicious implementations in daily life and pharmaceutical industries. Moreover, there is a distinct lack in utilizing the potential benefits of bioactive compounds from mushroom unless in vivo and in vitro studies are demonstrated.

Solid-State Fermentation (SSF) versus Submerged Fermentation (SmF) for the Recovery of Cellulases from Coffee Husks: A Life Cycle Assessment (LCA) Based Comparison

This article studies the environmental impacts of cellulase production by using a comparative attributional life cycle assessment (LCA) of two different scenarios of production. The first one is the commonly used submerged fermentation (SmF) using a pure substrate (cellulose powder) and a specific microorganism (Trichoderma reesei). The second scenario considers a novel system to produce enzymes and simultaneously treat a waste using the solid-state fermentation (SSF) process of coffee husk (CH) used as substrate. Experimental data were used in this scenario. The complete production process was studied for these two technologies including the fermentation phase and the complete downstream of cellulase. Life cycle inventory (LCI) data were collected from the database EcoInvent v3 (SimaPro 8.5) modified by data from literature and pilot scale experiments. The environmental impacts of both production systems revealed that those of SmF were higher than those of SSF. A sensitivity analysis showed that the results are highly conditioned by the energy use in the form of electricity during lyophilization, which is needed in both technologies. The results point to a possible alternative to produce the cellulase enzyme while reducing environmental impacts.

Statistical Optimization of the Physico-Chemical Parameters for Pigment Production in Submerged Fermentation of Talaromyces albobiverticillius 30548.

Talaromyces albobiverticillius 30548 is a marine-derived pigment producing filamentous fungus, isolated from the La Réunion island, in the Indian Ocean. The objective of this study was to examine and optimize the submerged fermentation (SmF) process parameters such as initial pH (4–9), temperature (21–27 °C), agitation speed (100–200 rpm), and fermentation time (0–336 h), for maximum production of pigments (orange and red) and biomass, using the Box–Behnken Experimental Design and Response Surface Modeling (BBED and RSM). This methodology allowed consideration of multifactorial interactions between a set of parameters. Experiments were carried out based on the BBED using 250 mL shake flasks, with a 100 mL working volume of potato dextrose broth (PDB). From the experimental data, mathematical models were developed to predict the pigments and biomass yields. The individual and interactive effects of the process variables on the responses were also investigated (RSM). The optimal conditions for maximum production of pigments and biomass were derived by the numerical optimization method, as follows—initial pH of 6.4, temperature of 24 °C, agitation speed of 164 rpm, and fermentation time of 149 h, respectively.

Solid-state fermentation increases secretome complexity in Aspergillus brasiliensis

Aspergillus is used for the industrial production of enzymes and organic acids, mainly by submerged fermentation (SmF). However, solid-state fermentation (SSF) offers several advantages over SmF. Although differences related to lower catabolite repression and substrate inhibition, as well as higher extracellular enzyme production in SSF compared to SmF have been shown, the mechanisms undelaying such differences are still unknown. To explain some differences among SSF and SmF, the secretome of Aspergillus brasiliensis obtained from cultures in a homogeneous physiological state with high glucose concentrations was analyzed. Of the regulated proteins produced by SmF, 74% were downregulated by increasing the glucose concentration, whereas all those produced by SSF were upregulated. The most abundant and upregulated protein found in SSF was the transaldolase, which could perform a moonlighting function in fungal adhesion to the solid support. This study evidenced that SSF: (i) improves the kinetic parameters in relation to SmF, (ii) prevents the catabolite repression, (iii) increases the branching level of hyphae and oxidative metabolism, as well as the concentration and diversity of secreted proteins, and (iv) favors the secretion of typically intracellular proteins that could be involved in fungal adhesion. All these differences can be related to the fact that molds are more specialized to growth in solid materials because they mimic their natural habitat.

Comparative Evaluation on Tannase Production by Lasiodiplodia plurivora ACN-10 under Submerged Fermentation (SmF) and Solid State Fermentation (SSF)

Aim: Tannase (tannin acyl hydrolase, E.C. 3.1.1.20) catalyzes the hydrolysis of ester bonds from complex hydrolysable tannins with the production of gallic acid and glucose and possess broad applications in biotechnology. This study is aimed at the production of tannase by Lasiodiplodia plurivora ACN-10 in SmF and SSF using Terminalia cattapa (almond leaves) and Magnifera indica (mango leaves) as substrates.&#x0D; Study Design: The design adopted to evaluate the production of tannase is submerged (SmF) and solid state (SSF) fermentation.&#x0D; Study Area: Federal Institute of Industrial Research Oshodi, Lagos State Nigeria.&#x0D; Methodology: Fifteen different soil samples were indiscriminately collected within Oshodi, Lagos, Nigeria and were inoculated on PDA plates at 30°C for 3-5 days. A total of 30 isolates was screened on Czapek dox minimal agar incorporated with 1% tannic acid and plates were incubated for 96 h at 30°C. Fungal isolates which were able to disintegrate tannic acid produced a clear halo zone around the colony diameter and were selected to be positive for tannase activity. The best isolate was identified based on its morphological, microscopic and molecular characteristics. Thereafter production and extraction of tannase was carried out in SmF for 0-120 h and in SSF for 0-144 h using Terminalia cattapa (Almond leaves) and Magnifera indica (Mango leaves) as substrates.&#x0D; Results: The total fungal count ranged from 1.0×104 to 3.5×105 CFU/g. A total of 30 fungal isolates produced clear halo zones (ranging from 20 to 70 mm) around the colonies during the screening with tannic acid. Isolate ACN-10, which showed the highest tannic degradation was identified based on its morphological and microscopic characteristics. On the basis of 18S rRNA gene sequence studies, the isolate was identified as Lasiodiplodia plurivora strain ACN-10 and the sequence was submitted to the Genbank with the accession number: MG250374. Results obtained in this study indicated that both substrates can be used by the isolate for tannase production in both SmF and SSF. The result of our investigation on the use of Terminalia cattapa (almond leaves) and Magnifera indica (mango leaves) as substrates for tannase production showed that optimum yield (6.064 U/ml) was obtained at 120 h in SSF while optimum production (4.623 U/ml) was observed in SmF at 96 h using Terminalia cattapa as substrate.&#x0D; Conclusion: Results obtained from this study indicated higher tannase production in solid-state fermentation compared to submerged fermentation. This is the first report to the best of our knowledge that Lasiodiplodia plurivora strain is implicated in tannase secretion. The result also demonstrated high production of extracellular tannases from low cost substrates which can be optimized and scaled up for industrial processes.

Comprehensive Optimization of Culture Conditions for Production of Biomass-Hydrolyzing Enzymes of Trichoderma SG2 in Submerged and Solid-State Fermentation.

Lignocellulose biomass contain large macromolecules especially cellulose and hemicelluloses that can be converted to fuel and chemicals using microbial biocatalysts. This study presents comprehensive optimization of production of biomass-hydrolyzing enzymes (BHE) by a high β-glucosidase-producing Trichoderma SG2 for bioconversion of lignocellulose biomass. Overall, a mixture of paper powder and switchgrass was most suited for production of BHE in submerged fermentation (SmF). BHE production was significantly different for various organic and inorganic nitrogen sources. The combination of peptone, yeast extract, and ammonium sulfate resulted in the highest activities (Units/mL) of BHE: 9.85 ± 0.55 cellulase, 38.91 ± 0.31 xylanase, 21.19 ± 1.35 β-glucosidase, and 7.63 ± 0.31 β-xylosidase. Surfactants comparably enhanced BHE production. The highest cellulase activity (4.86 ± 0.55) was at 25°C, whereas 35°C supported the highest activities of xylanase, β-glucosidase, and β-xylosidase. A broad initial culture pH (4-7) supported BHE production. The Topt for cellulase and xylanase was 50°C. β-xylosidase and β-glucosidase were optimally active at 40 and 70°C, respectively; pH 5 resulted in highest cellulase, β-glucosidase, and β-xylosidase activities; and pH 6 resulted in highest xylanase activity. Response surface methodology (RSM) was used to optimize major medium ingredients. BHE activities were several orders of magnitude higher in solid-state fermentation (SSF) than in SmF. Therefore, SSF can be deployed for one-step production of complete mixture of Trichoderma SG2 BHE for bioconversion of biomass to saccharide feedstock.

Optimization for bio-processing of elephant foot yam (Amorphophallus paeoniifolius) into Lacto-pickle using Taguchi statistical approach

Elephant foot yam (EFY) Lacto-pickles are crisp, tangy and are prepared by Lacto-fermentation. The study investigated the production of EFY Lacto-pickles in submerged fermentation (SmF) from EFY tubers using Lactobacillus (Lb) plantarum (MTCC-1325) as a controlled starter culture. Taguchi statistical method was used to estimate the effects of three optimum variables, [i.e., salt concentration (NaCl 8%, w/v), inoculum volume (10%, v/v), and incubation period (22 days)] on lactic acid (LA) production and shelf-life (SL) of Lacto-pickles using an L9 orthogonal array (OA) analysis. The optimum setting of the predicted value for LA production (i.e., 3.69289 g/kg) and SL of EFY Lacto-pickles (42 days) was found very approaching to the experimental value [LA (i.e., 3.873 g/kg) and SL (42 days)] supporting the validity of the technique. A significant reduction in hardness (215.0–179.0 N) and change in colour was attained with an increase in storage/shelf-life (42 days) (p ≤ 0.05) of EFY Lacto-pickles.

Polygalacturonase by Aspergillus Niger Using Seaweed Waste Under Submerged Fermentation: Production, Purification and Characterization

Crude oil, originating from fossilized organic materials, constitutes a crucial financial resource for all the countries with established deposits [1,2]. In general, oil reserves are allocated on the shelves of the ocean, on the land, as well as in the inland seas....

The amylase production by Actinobacteria isolated from rumen fluid

Amylolytic actinobacteria were isolated from the rumen fluid of ruminants. Eighteen isolates actinobacteria were obtained and the generation of actinobacteria mutants using chemical agent Sodium Azide. The actinobacteria wild type and mutants were then screened for amylase activity using starch agar plate assay. Ten isolates showed the ability to produce amylase enzymes. Submerged fermentation (SmF) was used for microbial production of amylase. It was found that maximum amylase activity (2.53 U/mL) was produced by actinobacteria isolate R-9 M, and 2.43 U/mL was produced by isolate R-2 WT using pollard as a substrate in Submerged fermentation. This preliminary study could provide base information for the discovery of novel actinobacteria from natural resources such as rumen fluid for the production of amylase, which will be used for multipurpose.