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陈雅娟,董园,芦志龙,陈东,黄日波.碱性纤维素酶产生菌株的筛选及其酶学性质研究[J].广西科学,2014,21(1):28-33. [点击复制]
- CHEN Ya-juan,DONG Yuan,LU Zhi-long,CHEN Dong,HUANG Ri-bo.Screening of Alkaline Cellulase-producing Strains and Research on Enzymatic Characteristics[J].Guangxi Sciences,2014,21(1):28-33. [点击复制]
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| 碱性纤维素酶产生菌株的筛选及其酶学性质研究 |
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陈雅娟1, 董园1, 芦志龙2, 陈东2, 黄日波1,2
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| (1.广西大学生命科学与技术学院, 广西南宁 530004;2.广西科学院, 非粮生物质酶解国家重点实验室, 国家非粮生物质能源工程技术研究中心, 广西生物质产业化工程院, 广西生物炼制重点实验室, 广西南宁 530007) |
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| 摘要: |
| [目的]从堆肥中筛选和鉴定产碱性纤维素酶的菌株,对菌株及其酶学特性进行研究。[方法]以刚果红平板染色法进行初筛,摇瓶发酵复筛,分离到高酶活菌株C73。通过形态观察和16S rRNA序列分析对菌株进行鉴定。采用DNS定糖法测定发酵液酶活最适pH值、pH值稳定性、最适温度、热稳定性及常见金属离子、SDS、EDTA等对酶活的影响。[结果]该菌株形态呈杆状,革兰氏阳性,不产芽孢,适宜在pH值8.0~11.0、30~37℃条件下生长。16S rRNA序列比对发现,该菌株与琼斯氏菌(Jonesia sp.)PC1序列相似度大于99%,初步鉴定为琼斯氏菌属菌株。在发酵培养基中,菌体浓度和CMCase活力分别在40h和50h达到最大值,随着发酵时间的延长发酵液pH值稳定在9.5左右。粗酶液的最适作用pH值为10.0,此时CMCase活力达2.3U/mL。在pH值8.0~11.0的范围内保持60%以上的酶活;最适作用温度为60℃,且在30~80℃的范围内保持60%以上的酶活。金属离子Mn2+、Co2+和(NH4)2SO4对酶活力有较强的抑制作用。[结论]菌株C73呈现较高的碱性CMCase酶活、较高的热稳定性和pH值稳定性,具有良好的工业应用前景。 |
| 关键词: 纤维素酶 琼斯氏菌 酶学性质 |
| DOI： |
| 投稿时间：2013-04-02修订日期：2013-10-12 |
| 基金项目:国家973项目(2010CB736209),国家863项目(2012AA022106),国家国际合作项目(2010DFB63590,2011DFA61910),广西科学研究与技术开发计划项目(桂科合10100019-21,桂科攻1099071,桂科合1140010-15),广西自然科学基金项目(2012GXNSFAA053062),广西科学院基本科研业务费项目(11YJ24SW08),广西科技创新能力与条件建设计划项目(桂科能12237022),广西科学基金项目(13-051-08)资助。 |
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| Screening of Alkaline Cellulase-producing Strains and Research on Enzymatic Characteristics |
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CHEN Ya-juan1, DONG Yuan1, LU Zhi-long2, CHEN Dong2, HUANG Ri-bo1,2
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| (1.College of Life Science & Technology of Guangxi University, Nanning, Guangxi, 530004, China;2.Guangxi Academy of Sciences, State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Industrialization Engineering Institute, Guangxi Key Laboratory of Biorefinery, Nanning, Guangxi, 530007, China) |
| Abstract: |
| [Objective] Strain with alkaline CMCase-producing ablities was screened from compost heaps and identified.Studies were conducted on the strain and its enzymatic characters.[Method] Strain C73 was screened primarily through Congo-red agar plates and secondly by flask fermentation.The strain was identified by morphism speculation and 16S rRNA sequence analysis.Enzymatic properties of optimal activity pH, pH stablity, optimal activity temperature and thermo-stablity as well as influence of metal ions, SDS and EDTA on the enzyme were determined throuth DNS method.[Result] The strain was rod-shaped, gram-positive, non-sporulating and could grow at pH 8.0~11.0, 30~37℃.Fermentation with CMC media showed maxium cell growth and CMCase avtivity peaks were reached at 40h and 50h, with the pH of system decreased to 9.5.Optimal CMCase activity pH was 10.0, while the CMCase activity reached 2.3U/mL.60% of CMCase activity was obtained at the pH range of 8.0~11.0.Optimal CMCase activity temperature was 60℃ and 60% of residual CMCase activity was obtained after incubation at 30~80℃ for 1 h.The addition of Mn2+, Co2+ and (NH4)2SO4 strongly inhibited CMCase activities.[Conclusion] Strain C73 exhibited higher alkaline CMCase activity, higher pH stablity and thermo-stablity and had a potential in industrial application. |
| Key words: cellulase Jonesia sp. enzymatic properties |
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