Optimization of procedural factors for advanced xylanase synthesis by Lysinibacillus fusiformis using Kolanut husk as substrate (original) (raw)
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Paddy Husk as Support for Solid State Fermentation to Produce Xylanase from Bacillus pumilus
Rice Science, 2011
To optimize culture conditions for xylanase production by solid state fermentation (SSF) using Bacillus pumilus, with paddy husk as support, solid medium contained 200 g of paddy husk with 800 mL of liquid fermentation medium [xylan, 20.0 g/L; peptone, 2.0 g/L; yeast extract, 2.5 g/L; K2HPO4, 2.5 g/L; KH2PO4, 1.0 g/L; NaCl, 0.1 g/L; (NH4)2SO4, 2.0 g/L, CaCl2·2H2O, 0.005 g/L; MgCl2·6H2O, 0.005 g/L; and FeCl3, 0.005 g/L] at pH 9.0 was applied. The highest xylanase activity (142.0 ±0.47 U/g DM] was obtained on the 6 th day at 30°C. The optimized paddy husk to liquid fermentation medium ratio was 2:9, and the optimized culture temperature was 40°C. When commercial Birchwood xylan was replaced with different concentrations of corncob, xylanase production was maximized (224.2 U/g DM) in the medium with 150 g/L corncob. Xylanase production was increased by sucrose, fructose and arabinose, whereas reduced by glucose, galactose, lactose and amylose. When organic nitrogen sources were replaced with locally available nitrogen sources such as groundnut powder or sesame seedcake powder or coconut seedcake powder or soy meal powder, the highest xylanase production (290.7 U/g DM) was obtained in the medium with soy meal powder and 16.0 g/L of soy meal powder was the optimum (326.5±0.34 U/g DM). Based on the optimization studies, B. pumilus produced 2.3 times higher xylanase activity.
Value addition of corn husks through enzymatic production of xylooligosaccharides
Brazilian Archives of Biology and Technology, 2016
Corn husks are the major wastes of corn industries with meagre economic significance. The present study was planned for value addition of corn husk through extraction of xylan, followed by its enzymatic hydrolysis into xylooligosaccharides, a pentose based prebiotic. Compositional analysis of corn husks revealed neutral detergent fibre 68.87%, acid detergent fibre 31.48%, hemicelluloses 37.39%, cellulose 29.07% and crude protein 2.68%. Irrespective of the extraction conditions, sodium hydroxide was found to be more effective in maximizing the yield of xylan from corn husks than potassium hydroxide (84% vs. 66%). Application of xylanase over the xylan of corn husks resulted into production of xylooligosaccharides with different degree of polymerization namely, xylobiose and xylotriose in addition to xylose monomer. On the basis of response surface model analysis, the maximum yield of xylobiose (1.9 mg/ml) was achieved with the enzymatic hydrolysis conditions of pH 5.8, temperature 44°C, enzyme dose 5.7U/ml and hydrolysis time of 17.5h. Therefore, the corn husks could be used as raw material for xylan extraction vis a vis its translation into prebiotic xylooligosaccharides.
3 biotech, 2017
Growth of Fusarium sp. BVKT R2, a potential isolate of forest soils of Eastern Ghats on birchwood xylan in mineral salts medium (MSM) under un-optimized conditions of 30°C, pH of 5.0, 150 rpm and inoculum size of 5 agar plugs for 7 days, yielded titer of 1290 U/mL of xylanase (EC 3.2.1.8). The effect of various operating parameters such as different substrates and their concentration, additional carbon and nitrogen sources, incubation temperature, initial pH, agitation and inoculum size on the production of xylanase by Fusarium sp. BVKT R2 was studied in shake flask culture by one factor at a time approach. The same culture exhibited higher production of xylanase (4200 U/mL) when grown on birch wood xylan in MSM under optimized conditions with an additional carbon source-sorbitol (1.5%) nitrogen source-yeast extract (1.5%) temperature of 30°C, pH of 5.0, agitation of 200 rpm and inoculum of 6 agar plugs for only 5 days. There was enhancement in xylanase production under optimized conditions by 3.2 folds over yields under unoptimized conditions. Growth of BVKT R2 culture on locally available lignocelluloses-sawdust, rice straw and cotton stalk-in MSM for 5 days released soluble sugars to the maximum extent of 52.76% with respect to sawdust indicating its greater importance in saccharification essential for biotechnological applications.
Advances in Enzyme Research, 2016
Xylanases are mostly produced through submerged fermentation; nonetheless solid-state fermentation has increased profound attention and consideration of scholars having high conversion level biomass to energy conservation. This study depicted the purification of xylanases and their possible utilization in industry. The present study was carried out to examine the culture influence of fungal strain Fomes fomentarius (F. fomentarius) using different agro-industrial residues (wheat straw, rice husk, sugarcane bagasse and siris pods). F. fomentarius showed maximum enzyme production after 72 h of fermentation, when grown on wheat straw in solid state fermentation process while maximum activity showed on pH 6.0 at 30˚C. The other parameters optimized by statistical design (RSM) showed maximum xylanase activity (146 ± 8 IU/mL) at 65% moisture content, 4 mL inoculums size, 175 mg Ammonium sulphate, 200 mg Calcium carbonate and 1.4 grams of glucose. Xylanase was salted out at 60% ammonium sulphate concentration and enzyme was further purified by Sephadex G-100 gel filtration chromatography with 2.2 fold increase in activity. The purified xylanase from F. fomentarius had optimum pH 6.0 and 40˚C. Xylanase showed higher specificity for oat spelt xylan with kinetic constants Km 1.25 mg/mL and Vmax 54 mM/min. Xylanases have an industrial important enzyme used extensively in food, feed and paper industry.
Sawdust and digestive bran as cheap alternate substrates for xylanase production
African Journal of …, 2011
Xylanases are a major group of enzymes, mostly produced from microbial fermentation processes, and have wide industrial and biotechnological applications. The production cost of xylanase is the major factor limiting its use, thus indicating the need for low cost production systems for market of this enzyme. In this study, therefore, sawdust and digestive bran were investigated as substrates for xylanase production by Bacillus strains. The xylanase titre ranging between 30.849 to 45.206 nkat/ml and 6.633 to 22.717 nkat/ml was produced by these Bacillus strains, using sawdust and wheat bran as the substrate, respectively. The optimum temperature for the production of xylanase was found to be between 45 and 55°C, while the optimum pH was 8.0 for all the strains tested. The xylanases produced by these Bacillus strains were found to be stable over a wide range of temperature tested (40 to 90°C). Up to 98 and 95% of the initial activity was retained by the crude extract of two of the Bacillus strains tested, while 10.3 to 56% loss in activity was observed for the other isolates after one hour incubation at 70°C. Addition of metal salts or additives to the crude extract was found to inhibit the enzyme activity to a varying degree, with the following order Hg 2+ > EDTA > Na > Urea > Mg 2+ > Ca 2+ observed. Findings from this study indicate the potential use of sawdust and digestive bran as cheaper alternatives for the production of xylanases.
Crude Xylanase Production in Bacteria using Corn Husk as Substrate in Submerged Fermentation
NU. International Journal of Science, 2019
Management of crop waste is important for sustainable agriculture. We investigated xylanase producing strains of local bacteria using corn husk as a substrate in submerged fermentation. Soil samples were collected from corn silage waste areas in Mae Sai, Rong Kwang District, Phrae Province, Thailand. Primary screening of crude xylanase producing isolates was made with the enrichment method in Mineral Salt Medium (MSM) containing 1% (w/v) corn husk on a shaking incubator at 150 rpm at 37oC for 7 days. In order to select potential isolates for further study, the serial dilution method was employed using MSM solid agar supplemented with xylan 0.1% (w/v). The enrichment method gave 236 bacterial isolates and these were used to determine their crude xylanase activity in spot diffusion and gel diffusion assays. Eleven isolates presented clear zones between 30 - 35 mm in diameter. The quantity of reducing sugars released in the supernatant in submerged fermentation was measured with the al...
Enzyme Research, 2011
A xylanase producer Bacillus mojavensis strain, called AG137, isolated from cotton farm (Kashan-Iran). The optimal xylanase activity reached at 55 • C & pH 9.0. Enzyme yield was studied using a medium with different agricultural wastes as inducers. Xylanase production of about 249.308 IU/mL was achieved at pH 8 and 37 • C, within 48 h submerged fermentation in enzyme production medium supplemented with 2% (w/v) oat bran as an optimum carbon source. A mixture of 1% (w/v) yeast extract and 1% (w/v) tryptone as optimum nitrogen sources, agitation speed 200 rpm, and inoculum size 2% (v/v) were the optimums for maximum production. Accordingly, xylanase yield from 194.68 IU/mL under non-optimized fermentation condition enhanced to 302.466 IU/mL in optimized condition. Screened xylanase is thermostable, presenting 70% stability at 60 • C during 30 min. Further enzyme incubation in higher temperature caused a decrease in the residual enzyme activity, yet it retained 68%-50% of its activity after 1 hour from 45 • C to 55 • C. Besides, it is stable in pH 9 and 10, maintaining over 70% of its activity for 2 h. The enzyme also could preserve 71% and 63% of its initial activity after 3 hours of pre-incubation in the same alkaline condition. Produced xylanase therefore was introduced as an alkaline-active and stable one, displaying suitable thermostability feature, confirmed by HPLC analysis. Hence, all xylanase properties highlight its promising uses in industrial scale.
ACS Omega, 2020
Over the past two decades, birchwood and beechwood xylans have been used as a popular substrate for the characterization of xylanases. Recently, major companies have discontinued their commercial production. Therefore, there is a need to find an alternative to these substrates. Xylan extraction from Acacia sawdust resulted in 23.5% (w/w) yield. The extracted xylan is composed of xylose and glucuronic acid residues in a molar ratio of 6:1 with a molecular mass of ∼70 kDa. The specific optical rotation analysis of extracted xylan displayed that it is composed of the D-form of xylose and glucuronic acid monomeric sugars. The nuclear magnetic resonance analysis of extracted xylan revealed that the xylan backbone is substituted with 4-O-methyl glucuronic acid at the O2 position. Fourier transform infrared analysis confirmed the absence of lignin contamination in the extracted xylan. Xylanase from Clostridium thermocellum displayed the enzyme activity of 1761 U/mg against extracted xylan, and the corresponding activity against beechwood xylan was 1556 U/mg, which confirmed that the extracted xylan could be used as an alternative substrate for the characterization of xylanases.