蝇蛆甲壳素对深绿木霉T2产生几丁质酶的诱导作用

The induction effects of flyblow chitin on chitinase production of Trichoderma atroviride T2

分别采用EDTA除钙和氢氧化钠-戊醇反应体系,并结合微波加热处理2次的优化工艺,从蝇蛆中提取甲壳素、壳聚糖,探讨二者对深绿木霉T2菌株产生几丁质酶的影响,并对深绿木霉T2几丁质酶活性及其对4种病原真菌的拮抗作用进行了研究。结果表明,提取的蝇蛆甲壳素的产出率为75.0%,灰分含量为17.3%;蝇蛆壳聚糖制备获得脱乙酰度为95.5%、含水量为8.36%的高质量壳聚糖产品,且碱用量明显下降,反应时间由4 h缩短为6m in;蝇蛆甲壳素培养基诱导培养的深绿木霉T2几丁质酶活性为0.9721U/mL,高于PDB液体培养基、SCMS营养液和蝇蛆壳聚糖培养基;不同营养条件下,蝇蛆甲壳素培养基均能够诱导深绿木霉T2菌株产生几丁质酶,且最佳产酶培养时间为120 h,酶活性峰值为1.8340U/mL;经蝇蛆甲壳素诱导后的深绿木霉T2菌株对4种病原菌的抑菌效果明显高于对照菌株T2。

The high quality flyblow chitin and flyblow chitosan were extracted from flyblow using optimizing processes of calcium removal of EDTA and reaction system of sodium hydroxide-pentanol with two times microwave heating respectively,was used to induce Trichoderma atroviride T2 produced more chitinase.The activity of chitinase by induced T.atroviride T2 were also determined and the antagonistic effects of induced T.atroviride T2 on four pathogenic fungi were detected.The results showed that the output ratio of flyblow chitin and the ash content extracted by using this optimized technology reached 75.0% and 17.3%.The high quality chitosan which contained 95.5% deacetylating degree of chitosan and 8.36% water content were obtained.Sodium hydroxide dosage was used less and reaction time shortened from 4 h to 6 min.The chitinase activity of T.atroviride T2 cultured in flyblow chitin medium was obviously higher than others cultured in PDB,SCMC and chitosan mediums,which were 0.9721 U/mL,0.4565 U/mL,0.7665 U/mL and 0.7315 U/mL,respectively.In different mediums,the optimum time for chitinase production in T.atroviride T2 was 120 h and the highest activity of chitinase was 1.8340 U/mL.The inhibition effects of T.atroviride T2 cultured in flyblow chitin medium antagonized on four pathogenic fungi,were higher than control.