| 摘要: |
| [目的]通过培养白腐菌Trametes pubescens提取胞外漆酶.[方法]以白腐菌T.pubescens MB89为漆酶培养菌,在25℃120 r/min条件下用基础GP液体培养基(300 mL培养基中含40 g/L丙三醇、15 g/L蛋白胨和1 g/LMgSO4·7H2O)进行胞外漆酶的震荡培养,培养10 d后,添加CuSO4·5H2O使培养基中Cu2+ 的最终浓度达1 mmol/L.定时测定漆酶活力,于漆酶活力开始下降时终止培养.将菌体破碎、离心后,用DEAE-Ssepharose对获得的漆酶进行分离和纯化.[结果]在以葡萄糖为碳源的培养基中添加一定浓度的Cu2+ ,可促进菌体的生长和漆酶活力的增加.培养前10 d,漆酶活力增加缓慢,培养10 d后添加CuSO4·5H2O,漆酶活力快速增加,并于培养的第19天达到最大,为51.70 U/mL.用超滤方法分离纯化漆酶,可使漆酶纯度达53%.[结论]T.pubescens MB89可作为生产漆酶的菌种,在漆酶培养过程中,Cu2+对漆酶活力的增加有促进作用. |
| 关键词: 漆酶 白腐菌 培养与纯化 |
| DOI： |
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| 基金项目:2004年奥地利政府项目 |
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| Formation and purification of laccase by the white-rot fungus Trametes pubescens |
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| Abstract: |
| 【Objcetive】 The study was about laccase formation and purification by white-rot fungus Trametes pubescens MB 89.【Method】The white-rot fungus Trametes pubescens MB 89 had been identified as an excellent producer of the industrially important enzyme laccase.Extracellular laccase formation could be considerably stimulated by the addition of Cu(Ⅱ) in the millimolar range to a simple,glucose-based culture medium.In this study,the final concentration of copper was 1 mmol/L in the medium,which was added CuSO4·5H2O after 10 d cultivation.When using a basal GP (glycerol-peptone) medium for cultures contained 40 g/L glycerol,15 g/L peptone from meat and 1 g/L MgSO4·7H2O,significant laccase formation by T.pubescens only started when glucose was completely consumed from the culture medium.DEAE-Sepharose was used for laccase purification.【Result】 Laccase activity increased slowly during the 10 d cultivation.Maximal laccase activiy obtained in shake-flask cultures was approx 57.10 U/mL on the 19thd.Purity of the laccase was above 53% by using DEAE-Sepharose.【Conclusion】 T.pubescens MB89 could be the productive strain of laccase in industry.Cu2+ was available for laccase formation. |
| Key words: laccase Trametes pubescens culturation and purification |
