CAJ | 학술논문

对一株高温蛋白酶高产菌株枯草芽孢杆菌BY25的发酵培养基组成与产酶条件进行了优化。正交试验结果显示培养基中各因子对产酶影响从高到低为：豆饼粉、葡萄糖、硫酸镁、麸皮、磷酸氢二钠、氯化钙。在此基础上进行培养基组成优化，将豆饼粉、麸皮混合氮源改为以豆饼粉为单一氮源进行蛋白酶发酵。单因素试验发现，在单一氮源培养条件下，培养基中各因子对产酶影响从高到低依次为：豆饼粉、葡萄糖、氯化钙、磷酸氢二钠。除微量氯化钙外，金属盐，尤其是金属硫酸盐的添加对产酶有显著抑制作用。此外，培养初始pH和培养时间对产酶有显著影响，接种量也有一定影响。通过绘制120 h产酶曲线发现，BY25产酶曲线为双峰，产酶曲线顶峰出现在发酵后72 h，优化后BY25发酵培养基各组分添加量为豆饼粉60 g/L、葡萄糖60 g/L、氯化钙0．5 g/L、磷酸氢二钠4 g/L，接种量为4．5％～5％，初始培养pH为8．0。优化产酶培养基和产酶条件后，发酵液酶活力可达到101．1μmol/（ min·mL）。
대일주고온단백매고산균주고초아포간균BY25적발효배양기조성여산매조건진행료우화。정교시험결과현시배양기중각인자대산매영향종고도저위：두병분、포도당、류산미、부피、린산경이납、록화개。재차기출상진행배양기조성우화，장두병분、부피혼합담원개위이두병분위단일담원진행단백매발효。단인소시험발현，재단일담원배양조건하，배양기중각인자대산매영향종고도저의차위：두병분、포도당、록화개、린산경이납。제미량록화개외，금속염，우기시금속류산염적첨가대산매유현저억제작용。차외，배양초시pH화배양시간대산매유현저영향，접충량야유일정영향。통과회제120 h산매곡선발현，BY25산매곡선위쌍봉，산매곡선정봉출현재발효후72 h，우화후BY25발효배양기각조분첨가량위두병분60 g/L、포도당60 g/L、록화개0．5 g/L、린산경이납4 g/L，접충량위4．5％～5％，초시배양pH위8．0。우화산매배양기화산매조건후，발효액매활력가체도101．1μmol/（ min·mL）。
The fermentation media components and culture conditions of the thermostable protease from a thermophilic strain Bacillus subtilis BY25 were further studied. A six factor and three level orthogonal experiment was conducted to ob-serve the effects of fermentation medium components on thermostable protease yield. Wheat bran was replaced by soybean cake powder for its low contribution rate. Single factor experiment was carried out to optimize the protease production. The optimized culture condition was at 30℃, with initiating pH of 8.0, 4.5%-5% inoculum size for 72 h;and the opti-mized fermentation medium components were described as follows:glucose 60 g/L, soybean cake powder 60 g/L, Na2 HPO4 4 g/L and CaCl2 0.5 g/L. After fermentation, the casein hydrolysis activity of the enzyme was estimated up to 101.1 μmol/( min·mL) , much higher than other domestic reports.