欢迎访问《草业学报》官方网站,今天是 分享到:

草业学报 ›› 2014, Vol. 23 ›› Issue (2): 243-252.DOI: 10.11686/cyxb20140229

• 研究论文 • 上一篇    下一篇

东祁连山高寒草地土壤产漆酶真菌的筛选、鉴定及产酶条件的初步研究

芦光新1,王军邦2*, 陈秀蓉3*,杨成德3,薛莉3   

  1. 1.青海大学农牧学院草业科学系,青海 西宁 810016;
    2.中国科学院地理科学与资源研究所,北京 100094;
    3.甘肃农业大学 草业学院 草业生态系统教育部重点实验室 中-美草地畜牧业可持续发展研究中心,甘肃 兰州 730070
  • 收稿日期:2013-05-23 出版日期:2014-02-25 发布日期:2014-04-20
  • 通讯作者: E-mail:jbwang@igsnrr.ac.cn,chenxiurong@gsau.edu.cn
  • 作者简介:芦光新(1974-),男,青海湟中人,教授,博士。E-mail:lugx74@qq.com
  • 基金资助:
    国家自然科学基金“青藏高原草地耐低温纤维素分解真菌多样性研究”(41261064)和青海大学高层次人才基金“产漆酶真菌的筛选及多酶系菌群的构建”(2012-QGC-9)资助。

Study on a laccase-producing fungus from alpine grassland soil in eastern Qilian Mountains: screening, identification, and activity analyses

LU Guang-xin1, WANG Jun-bang2, CHEN Xiu-rong3, YANG Cheng-de3, XUE Li3   

  1. 1.Department of Grassland Science, Agriculture and Animal Husbandry College, Qinghai University, Xining 810016, China;
    2.Institute of Geographic Sciences and Nature Resources Research, CAS, Beijing 100101, China;
    3.Pratacultural College, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2013-05-23 Online:2014-02-25 Published:2014-04-20

摘要: 以高寒草地土壤分离出的56个真菌菌株为研究对象,经愈创木酚-PDA培养基和α-萘酚-PDA选择性培养基初步筛选, 再根据菌株在愈创木酚、邻苯二酚、邻苯甲苯胺为底物的选择性培养基上菌体生长及菌落大小、漆酶催化氧化还原反应产生的变色圈直径大小及其变色圈颜色深浅程度,以及测定油菜秸秆诱导的液体发酵产漆酶活力,筛选出1株产漆酶酶活真菌菌株(编号为310b)。经rDNA-ITS基因序列分析,初步鉴定为Marasmius tricolor。对菌株310b产漆酶的条件进行了初步研究,结果表明,菌株310b在25℃培养条件下产漆酶活力最大,初始pH值为4.0时,漆酶活力最大,蔗糖、蛋白胨分别为诱导菌株产漆酶活力最高的碳、氮源。几种非营养有机物对菌株产漆酶活力大小不同,α-萘酚和吐温-80对菌株产漆酶没有明显影响,愈创木酚、单宁酸、吲哚乙酸抑制菌株产漆酶,其中吲哚乙酸抑制作用最强烈(P<0.01)。在Cu2+浓度为0.001~0.025 g/L范围内,随Cu2+增加,产漆酶活力增加,0.025 g/L时产酶活力最大。随着接种量的增加,诱导漆酶活力增加。在60~180 r/min的转速范围内,随着转速的增加,漆酶活力增加。油菜秸秆粉的量在0~1 g范围内,随着秸秆添加量的增加,漆酶活力增加;1~2 g范围内,随着秸秆添加量的增加,漆酶活力减小。

Abstract: The sixty-five fungi strains that were isolated from the alpine grassland soil in the eastern Qilian Mountains were researched as the object in this paper. The fungi initially selected through culturing on the medium of guaiacol-PDA and α-naphthol-PDA medium. The fungi with the function to produce laccases was screened (identified as No. 310b) based on the growth and colony sizes on the culture mediums of guaiacol, caechol, and o-benzyl aniline, the activities of redox reaction catalyzed by the laccases in these fungi grown, as well as the activity of laccases in these fungi cultured on the stems of oil rape. By rDNA-ITS sequence analysis, the fungus was identified as Marasmius tricolor. Then elementary study on the conditions under which laccases were produced by the fungus (No. 310b) was carried out. These results showed that the activity of laccases in the fungus reached the peak when it was cultured under the condition: 25℃, initial pH 4.0, sucrose and peptone that were used as carbon and nitrogen resources, respectively. Few non-trophic organic chemicals showed different effects on laccase activity in the fungus. α-naphthol and Tween-80 did not have significant effects on laccase activity in the fungus, but guaiacol, tannic acid, and indole acetic acid inhibited the activity of laccases, especiall indole acetic acid, which significantly inhibited laccase activity (P<0.01). When Cu2+ concentrations ranged from 0.001-0.025 g/L, laccase activity increased as the concentrations of Cu2+ increased, reaching the peak at 0.025 g/L. With the increase of inoculum sizes, laccase activity increased. Ranging from 60 to 180 r/min, laccase activity increased. When the amount of the powder of oil rape stems ranged from 0 to 1 g, laccase activity increased, but when the amount was higher than 1, the activity decreased.

中图分类号: