仿刺参消化道内产琼胶酶菌株的选育及培养条件优化  

Breeding and its optimal culture of bacterial and agarase-producing strain from digestive tracts in sea cucumber Apostichopus japonicus

作　　者：马悦欣[1] 安军[1] 刘双连[1] 徐高蓉[1] 于书渤[1] 

机构地区：[1]大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室,辽宁大连116023

出　　处：《大连水产学院学报》2007年第2期86-91,共6页Journal of Dalian Fisheries University

基　　金：辽宁省自然科学基金资助项目(20062129)

摘　　要：对仿刺参Apostichopus japonicus消化道内容物进行富集培养,以琼胶为惟一碳源进行平板初筛,摇瓶复筛,得到一株琼胶酶活性较高的菌株HF3。对该菌株进行紫外线诱变,获得突变菌株HF3-04,酶活力从43.0 U/mL增加到48.9 U/mL,提高了13.7%。通过正交试验,确定培养基的最佳组成为:琼胶3 g/L,硝酸铵1 g/L,磷酸氢二钠0.5 mmol/L,pH值8.0;最佳培养条件为:接种量为1%,250 mL三角瓶中发酵培养基的装液量为110 mL,摇床转速为150 r/m in,培养温度为25℃,培养时间为48 h。将培养基组成及培养条件优化后,菌株酶活力达到80.7 U/mL,比优化前提高了65.0%。A bacterial agarase- producing strain HF3 was isolated from digestive tracts in sea cucumber Aposticho- pus japonicus by enrichment culture, first - screened by agar plate containing agar as main carbon source and sec- ond - screened by shaking culture. The strain HE3 was treated with UV for screening of high agarase - producing mutant. The mutant HF3 -04 had an increase activity of agarase from 43.0 U/mL to 48.9 U/mL, 13.7% higher than the control, and the higher production of the agarase was continued thereafter for five generations. The optimal culture media for HF3 -04 strain producing agarase was determined by single factor test and orthogonal test and was found as the following： agar 3 g/L, NH4NO3 1 g/L, Na2HPO4 ·12H20 0.5 mmol/L, and pH 8.0. The optimal fermentation conditions were observed as the following： inoculum volume 1%, 110 mL medium in 250 mL Edenmeyer flask, shaking speed 150 r/min, culture temperature 25℃ and culture period 48 h. The HF3 - 04 had agarase activity of 80.7 U/mL, 65.0% higher when it was incubated in optimal fermentation conditions.

关 键 词：仿刺参 琼胶酶 紫外线诱变 培养条件优化 

分 类 号：Q935[生物学—微生物学]