高产漆酶真菌的筛选及产酶条件研究  

Screening of Fungal Strains with High Yield of Laccase and Studying on the Laccase-Producting Conditions

作　　者：林娟[1,2] 兰帅琼[1] 杨捷[1,2] 李仁宽[1,2] 叶秀云[1,2] 

机构地区：[1]福州大学生物科学与工程学院,福州350108 [2]酶高效表达国家工程实验室,福州350002

出　　处：《中国食品学报》2013年第8期110-116,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：福建省发改委生物产业技术开发专项资助;2013年海洋公益性行业科研专项资助(201305015)

摘　　要：目的:对筛选得到的漆酶高产菌株进行鉴定和产酶条件优化,以提高漆酶活力。方法:采用愈创木酚-PDA和RB亮蓝-PDA平板初筛,得到1株产漆酶能力较强的菌株HYB07,对其进行18S rDNA序列分析,并结合菌丝和菌落形态观察,鉴定HYB07为一色齿毛菌(Cerrena unicolor)。以高氮低碳高无机盐培养基(LM3)为基础培养基,对其中的碳源、氮源、诱导物和培养基初始pH值进行优化,并研究其发酵产酶条件(接种量、发酵温度、转速、装液量),得到HYB07菌株的最佳产酶条件是:葡萄糖2%,酒石酸铵1.5%(ABTS 0.05 mmol/L,培养基初始pH 5.0,接种量8%(体积分数),温度25℃,转速200 r/min,装液量60 mL/250 mL,发酵时间6 d。结论:在上述条件下发酵,漆酶最高活力达79 733 U/L,比采用LM3培养基(56 356 U/L)提高了41.5%。Cu2+对漆酶的合成影响显著,缺乏Cu2+时,漆酶活力下降了87.64%。Objective： The strain screened from nature was identified and its laccase-producing conditions were opti- mized to increase laccase activity. Method： A strain HYB07 with high yield of laccase was obtained by screening with PDA plates containing guaiacol and Remazol brilliant blue R, respectively. Strain HYB07 was identified as Cerrena uni- color via 18s rDNA sequence analysis and morphology observation. Based on high nitrogen and low carbon medium （LM3）, the carbon sources, nitrogen sources, inducer and initial pH were optimized. Furthermore, the fermentation con- ditions such as inoculum volume, temperature, shaking speed and liquid volume in flask were explored. The optimal lac- case-producing conditions were achieved： glucose 2%, Ammonium tartrate 1.5%, ABTS 0.05 mmol/L, initial pH 5.0, in- oculum volume 8%, temperature 25 ℃, shaking speed 200 r/min, liquid volume in flask 60 mL/250 mL, fermentation pe- riod 6 d. Conclusion： Under the optimal conditions, the maximum laccase activity was 79 733 U/L, 41.5% higher than fermented with LM3 medium. Moreover, laccase production was significantly affected by Cu2＋, and laccase activity was decreased by 87.64% absence of Cu2＋.

关 键 词：真菌 漆酶 筛选 产酶条件 

分 类 号：TQ925[轻工技术与工程—发酵工程]