几丁质降解酶产生菌的分离及其产酶条件的初步研究

从土壤中分离出两株几丁质降解能力较强的细菌,经初步鉴定,分别归于假单胞菌属和短杆菌属,命名为Pseudomonassp.ww和Brevibacteriumsp.yy.对这两菌株进行的液体摇瓶发酵试验表明,在肉汤培养基中,yy菌株16h进入稳定期,ww菌株20h进入稳定期,而在几丁质培养基中两菌株要培养36h才进入稳定期;在几丁质培养基中对酶活力的测定结果表明:yy菌株培养35h酶活达最高,为8.3U/mL,ww菌株培养30h时酶活最高,达13.3U/mL.用半量肉汤培养基摇瓶培养10h后加入0.2%胶体几丁质的补料分批培养方式,可使酶活高峰时间提前5h,且酶活力也有所增加.yy菌株在30h酶活达最高,为8.4U/mL,ww菌株在25h酶活达最高,为15.6U/mL,若加入的是2%的胶体几丁质,则对菌体,特别是对yy菌株的生长有抑制作用.实验还发现两菌株的几丁质培养物对金黄色葡萄球菌有明显的抑制作用,对大肠杆菌也有一定的抑制作用,而对黑曲霉和黑根霉几乎没有抑制作用.

培养条件对嗜热芽孢杆菌HU1合成几丁质降解酶及脱乙酰基酶的影响

[目的]研究培养条件对嗜热芽孢杆菌HU1产酶的影响,为工业化发酵生产酶及壳寡糖的制备提供依据。[方法]采用摇瓶培养方式,对嗜热芽孢杆菌HU1产几丁质降解酶及脱乙酰基酶的培养条件进行研究。[结果]最佳诱导碳源为粉末几丁质,其最适添加量为3.5%;最佳氮源为酵母粉,最适添加量为1.0%;初始pH值为6.0时,有利于产酶;Cu2+对酶的合成表现出明显的抑制作用;而Ca2+则能有效增加产酶能力。Mg2+及Zn2+对嗜热芽孢杆菌HU1 2种酶的合成无显著影响;培养72 h,产酶量达到最高。以粉末几丁质为底物,对粗酶液的水解产物进行了分析,其12 h的水解产物主要集中于分子量为六糖以下的壳聚糖。[结论]利用粗酶液制备小分子壳寡糖工艺流程简单,为后期进一步开发保健品、生物农药、饲料添加剂、食品添加剂等奠定了基础。

Isolation and Screening of Strains for Silkworm Puparium Chitin Degradation and the Optimization of Chitinase Production by Response Surface Methodology

[Objective] The aim of this study was to optimize the conditions of chitinase-produce strains.[Method] A kind of screened chitinase-produce strain G-254 was habituated cultured,and then the single factor experiment was carried out to explore the effects of different carbon source,nitrogen source and inorganic salt on the activity of produced chitinase;the response surface test was used to determine the optimal conditions for chitinase production.[Result] The optimal conditions for chitinase production were：8% of glucose,5% of beef extract and 0.07% of MgSO4,and the activity of chitinase reached the maximal value（6.86 U）under these conditions.[Conclusion] The study improved the activity of chitinase produced by strain G-254 and provided good foundation for industrial production.