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METHOD OF PRODUCING ALKALOTHERMOSTABLE XYLANASE FROM BACILLUS PUMIIUS STRAIN MK001 BY SOLID STATE FERMENTATION
METHOD OF PRODUCING ALKALOTHERMOSTABLE XYLANASE FROM BACILLUS PUMIIUS STRAIN MK001 BY SOLID STATE FERMENTATION
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机译：固态发酵法从芽孢杆菌MK001中制备耐高温木聚糖酶的方法
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摘要
This invention relates to a novel method for production of cellulase free xylanase, an enzyme from Bacillus pumilus strain MK001 from wheat bran by solid state fermentation comprising, Pretreatment of wheat bran to remove substantial portion of starch by sieving crude wheat bran particles through 200 μ mesh resulting uniform particle size 180-200 n, washing, drying at 50°C, Followed by steaming the bran at 121°C for 30min and then moistening with 3.5 litre of 0.2-0.4 N preferably 0.3 N NaoH at room temperature for 24hr. and finally washing and overnight drying at 50°C, Supplementing the pretreated wheat bran with mineral slat solution containing salts as (g/1) KH2 Po4 0.9-1.1 preferably 1.0, Nacl2.2-2.6 preferably 2.5; Mg So4, 7H2o, 0.9-1.1 preferably 0.1; (NH4) 2 S04, 0.9-1.1 preferably 1.0; Cac12 0.9-1.1 preferably 0.1 and folic acid 0.28-0.50% preferably 0.28% W/W, maintaining the bacterial cultivation pH value between 8.5 to about 9.2, preferably at 9.0 by using 10% W/V Na2 Co3, and temperature about 36°C to 39°C, preferably at 37°C, Preparing seed culture by streaking B. pumilus strain MK 001 stock culture on a petri plate containing xylan-Horikoshi agar pH 8.0 containing % w/v birch wood xylan 0.5, peptone 0.5, yeast extract 0.5, KH2 Po4 0.1 Mg So4, 7H20 0.01, incubating at 37°C for 24h followed by inoculation with a loopful of 24h old culture of B.pumilus strain MK 001 in a flask containing 20ml of Horikoshi medium containing % W/V glucose 0.5, peptone 0.5, yeast extract 0.5, KH2 Po4 0.1 Mg 804, 7H2O 0.01, pH 8.0 at 37°C under shaking conditions 200rpm for 6hr. Inoculating the enamel tray containing about 700.0 g of wheat bran with the culture medium as described in (iv) such that the ratio by volume of the culture medium to the nutrient medium in the enamel tray is conveniently 12.50 (v/w) to about 12.00, preferably 12.5, wherein the solid-state fermentation is carried out up to 120 hours. Extracting the enzyme from fermented wheat bran using 50mM-150mM preferably 100.0 m M citrate phosphate buffer pH 6.0 vortexing at 200-250rpm for 30 min at 37°C removing solid constituents by centrifuging at 10, 000 x g for 10 to 15 min at 4°C.
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