河北科技师范学院学报
河北科技師範學院學報
하북과기사범학원학보
JOURNAL OF HEBEI NORMAL UNIVERSITY OF SCIENCE & TECHNOLOGY
2011年
3期
58-63
,共6页
低温蛋白酶%筛选%发酵条件%酶活
低溫蛋白酶%篩選%髮酵條件%酶活
저온단백매%사선%발효조건%매활
cold-active protease%screening%fermentation condition%enzyme activity
采用平板初筛和摇瓶复筛等方法,从渤海秦皇岛海域的海水样品中筛选到产低温蛋白酶活力较高的菌株HYM-16,初步鉴定为黄色杆菌属,设计单因素和正交试验,确定该菌的最佳发酵条件。结果表明,在装液量50 mL液体培养基的250 mL三角瓶中,以体积分数0.01的接种量、摇床转速180 r/min条件下,25℃培养48 h酶活最高;最适碳源、氮源分别为蔗糖、豆饼粉,Mg2＋显著提高产酶量;采用L9（34）正交试验得最适发酵培养基配方：蔗糖1 g/L,豆饼粉20 g/L,MgCl2 0.4 g/L,K2HPO4 0.5 g/L,KH2PO4 0.5 g/L。经验证,优化后的发酵培养基菌株酶活达605.8×103 U.L-1,较基础培养基提高了46.9%。
採用平闆初篩和搖瓶複篩等方法,從渤海秦皇島海域的海水樣品中篩選到產低溫蛋白酶活力較高的菌株HYM-16,初步鑒定為黃色桿菌屬,設計單因素和正交試驗,確定該菌的最佳髮酵條件。結果錶明,在裝液量50 mL液體培養基的250 mL三角瓶中,以體積分數0.01的接種量、搖床轉速180 r/min條件下,25℃培養48 h酶活最高;最適碳源、氮源分彆為蔗糖、豆餅粉,Mg2＋顯著提高產酶量;採用L9（34）正交試驗得最適髮酵培養基配方：蔗糖1 g/L,豆餅粉20 g/L,MgCl2 0.4 g/L,K2HPO4 0.5 g/L,KH2PO4 0.5 g/L。經驗證,優化後的髮酵培養基菌株酶活達605.8×103 U.L-1,較基礎培養基提高瞭46.9%。
채용평판초사화요병복사등방법,종발해진황도해역적해수양품중사선도산저온단백매활력교고적균주HYM-16,초보감정위황색간균속,설계단인소화정교시험,학정해균적최가발효조건。결과표명,재장액량50 mL액체배양기적250 mL삼각병중,이체적분수0.01적접충량、요상전속180 r/min조건하,25℃배양48 h매활최고;최괄탄원、담원분별위자당、두병분,Mg2＋현저제고산매량;채용L9（34）정교시험득최괄발효배양기배방：자당1 g/L,두병분20 g/L,MgCl2 0.4 g/L,K2HPO4 0.5 g/L,KH2PO4 0.5 g/L。경험증,우화후적발효배양기균주매활체605.8×103 U.L-1,교기출배양기제고료46.9%。
Through preliminary petri-dish and following shaking flask fermentation screening,Strain HYM-16 producing higher protease was isolated from samples in Qinhuangdao of Bohai Sea located at 39°93′ N and 119°60′ E,and was primarily identified as Xanthobacter sp.Its fermentation condition was studied through the single-factor and orthogonal experiment.The results demonstrated that after 48 hours at 25 ℃ it had the maximum protease activity with 1% inoculation size and the stirring speed at 180 r/min,when 50 mL liquid culture in 250 mL flask.The optimal carbon and nitrogen source was saccharose and soybean flour respectively.Magnesium was beneficial to enzyme activity obviously.By using orthogonal experiment L9（34）,the top-seeded formula for the media（g/L） was obtained： saccharose 1,soybean flour 20,Mg2＋ 0.4,dipotassium hydrogen phosphate 0.5,and potassium dihydrogen phosphate 0.5.Finally,the optimized fermentation medium was validated,and the total activity of protesae was 605.8 U·mL-1,which was 46.9% higher than basic medium.
