N^＋注入选育漆酶高产菌株及其产酶优化研究  

SCREENING OF FUNGAL MUTANT STRAIN WITH HIGH LACCASE YIELD BY N^+-IMPLANTATION AND ENZYME PRODUCTION CONDITION OPTIMIZATION

作　　者：王力生[1] 朱洪龙[2] 李春凤[1] 程茂基[1] 蔡海莹[1] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]南京农业大学动物科技学院,江苏南京210095

出　　处：《核农学报》2009年第2期228-234,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金项目(30671526);安徽省科技攻关项目(06013040A)

摘　　要：以糙皮侧耳Pleurotus ostreatus WY01为出发菌株,通过N+注入诱变处理担孢子、RBBR-PDA平板变色法初筛、ATBS法测定漆酶活性复筛,获得1株漆酶高产诱变菌株ADW-08。用高碳低氮无机盐制备的油菜秸固体培养基(SM)培养,其峰值酶活力比出发菌株高出2.80倍,达7.78U/g,且产酶稳定。对ADW-08固体培养产酶条件的研究表明,以葡萄糖为碳源明显优于蔗糖、麦芽糖、麸皮和可溶性淀粉;酒石酸铵较有利于ADW-08漆酶的分泌;适宜初始pH为5.0或6.0;ABTS和藜芦醇对产酶均有明显的诱导作用,而吐温-80对产酶有一定的抑制作用;正交试验结果表明,葡萄糖、酒石酸铵和pH最佳参数分别为15.0g/L、0.2g/L和5.2,酶活峰值为8.33U/g。The basidiospores of Pleurotus ostreatus WY01 were exposed to N^＋ beam and the treated basidiospores were screened in RBBR-containing PDA plates. Then the laccase activities of the selected strains were determined by ABTS, and a mutant named ADW-08 with high yield of laccase was obtained. The maximum activity of laccase of ADW-08 increased up to 7.78 U/g, 2.8 times of the original stain, and its laccase production ability was stable in high carbon and low nitrogen rape straw solid medium （SM）. Results showed that glucose as carbon source was obviously superior to sucrose, maltose, wheat bran and soluble amylum in soild culture for ADW-08. Ammonium tartrate as nitrogen source was more suitable than other nitrogen sources for the laccase secretion. The optimal initial pH was 5.0 or 6.0. The ability of laccase production clearly increased by ABTS and veratryl alcohol induction, but was inhibited by Tween 80. When the optimum parameters for laccase production got from orthogonal design were as follows：glucose 15g/L, ammonium tartrate 0.2g/L,pH 5.2, the peak value of Lac activity was 8.33U/ g.

关 键 词：糙皮侧耳 N^+注入诱变 条件优化 漆酶 

分 类 号：TQ925[轻工技术与工程—发酵工程] TS745