CAJ | 학술논문

通过氮离子注入法诱变哈茨木霉(Trichoderma harzianum)野生菌,再以透明圈法对平板培养菌进行筛选,用DNS法测定液体培养菌的酶活,从中筛选出1株产几丁质酶能力高的哈茨木霉菌株H-13,并通过单因素试验和正交试验优化该目标菌株的发酵条件.单因素试验结果表明,添加1.0%蛋白胨时产酶量最高,1.0%胶体几丁质对菌体产几丁质酶的诱导性最强,初始pH 5.5、培养96 h时产酶量较高.正交试验表明,H-13以1.0%蛋白胨为氮源、0.5%胶体几丁质为碳源,在初始pH 5.0的培养基中培养96 h时,发酵液中的几丁质酶活可达到731.69 U·mL~(-1),比同样条件下的野生菌株酶活力提高1倍.
통과담리자주입법유변합자목매(Trichoderma harzianum)야생균,재이투명권법대평판배양균진행사선,용DNS법측정액체배양균적매활,종중사선출1주산궤정질매능력고적합자목매균주H-13,병통과단인소시험화정교시험우화해목표균주적발효조건.단인소시험결과표명,첨가1.0%단백동시산매량최고,1.0%효체궤정질대균체산궤정질매적유도성최강,초시pH 5.5、배양96 h시산매량교고.정교시험표명,H-13이1.0%단백동위담원、0.5%효체궤정질위탄원,재초시pH 5.0적배양기중배양96 h시,발효액중적궤정질매활가체도731.69 U·mL~(-1),비동양조건하적야생균주매활력제고1배.
Trichoderma harzianum wild-type strains were implanted by N~+ to produce mutagenesis. A mutant stain H-13 with higher activity of chitinase was obtained by screening the mutants with transparent zone, and analyzing chitinase activity by DNS method. Single-factor design and orthogonal test design were used for optimizing cultural conditions of chitinase production from mutant strain H-13. Single-factor experimentation showed that the media added 1.0% peptone led to highest chitinase production, and 1.0% colloidal chitin was the best media for inducing chitinase. The mutant strain showed higher enzymatic activity in this media with initial pH 5.5 for 96 h of culture. Orthogonal test showed that chitinase activity of strain H-13 could reach 731.69 U·mL~(-1) cultured in the optimal medium with 1.0% peptone, 0.5% colloidal chitin and initial pH 5.0 for 96 h, which was one fold increase compared to the wild strain enzyme vigor.