|
|
|
| Optimization of Fermentation Conditions of Collaborative Expression of Chitinase and Natamycin from Streptomyces lydicus A01-chit33 CT |
| LIN Zhen-ya, WU Qiong, SUN Rui-yan, CHEN Jie, LI Ya-qian |
| Key Laboratory of Urban Agriculture (South) of Ministry of Agriculture, School of Agriculture & Biology, Shanghai Jiaotong University, Shanghai 200240, China |
|
|
|
Abstract Biological control of plant diseases is an effective method with the beneficial microorganisms or microbial metabolites against crop diseases. Streptomyces are already identified to produce a range of antibiotics including natamycin etc., which exhibit an excellent activities against fungal pathogens. Streptomyces lydicus A01 has been proved as a biocontrol microbe with antagonistic activity against Botrytis cinerea by producing natamycin, which degrading the cell membrane of fungal pathogens. Natamycin, a polyene macrolide antibiotic with broad-spectrum antifungal activity, has a stable and strong inhibitory activity against many plant pathogenic fungi, and is commonly used as antifungal agent. Chitinase can degrade chitin and could be used for resist many fungal pathogens. In A01-chit33CT, natamycin was produced to inhibit plant pathogens via destructing fungal cell membrane, while chit33 plays an important role in hydrolyzing the chitin component in fungal pathogen cell wall. The coordination effect of natamycin and chit33 may be improved significantly.
The optimal fermentation conditions for chitinase activity and natamycin produced in Streptomyces lydicus A01-chit33CT were studied. Firstly, the effect of different carbon and nitrogen sources on natamycin production and chitinase activities were investigated. The results showed that: glucose can promote natamycin production but inhibit the expression of chit33, so glucose and chitin powder were added into the fermentation medium with two-stage method to achieve collaborative expressions of both. The optimal medium compositions were as follow: chitin powder 10 g/L(after 4 days), glucose 40 g/L, soybean meal 30 g/L,soya peptone 10 g/L,CaCO3 5g/L,MgSO4·7H2O 0.5g/L,K2HPO4 0.5 g/L. Secondly, the cultural conditions were further to optimum as follows: initial pH 6.0, temperature 28℃, rotation speed 180 r/min. The highest natamycin production could reach 1.52 g/L and chitinase activity reach 990U/ml under the optimal fermentation conditions. Both levels improved by 1.95 and 2.27 times at post-optimazation compared to that of pre-optimization, respectively.
|
|
Received: 29 June 2012
Published: 25 October 2012
|
|
|
|
|
[1] John L, Koont Z, Joseph E, et al. Formation of natamycin: cyclodextrin inclusion complexes and their characterization. Journal of Agricultural and Food Chemistry, 2003, 51: 7106-7110.
[2] Davidson P M, Doan C H. Natamycin. University of Idaho, 1993, 7:395-407.
[3] 李海波. 纳他霉素高产菌株的诱变育种.保定: 河北农业大学, 2008. Li H B. Studies on mutation and screening of strains producing natamycin. Baoding: Agricultural University of Hebei, 2008.
[4] 花玉鹏,文才艺,刘伟成,等. 响应曲面法优化利迪链霉菌A02发酵培养基. 中国生物防治学报, 2011, 27(4): 520-527. Hua Y P, Wen C Y, Liu W C, et al. Optimization of medium components for production of natamycin by Streptomyces lydicus A02 with response surface methodology. Chinese Journal of Biological Control, 2changes in perceptions derived from research on Trichoderma harzianum T-22. Plant Disease, 2000, 84: 377-393.
[6] 陈捷,朱杰伟,张婷,等. 木霉菌生物防治作用机理与应用研究进展. 中国生物防治学报, 2011, 27(2): 145-151. Chen J,Zhu J W,Zhang T, et al. Progress on mechanism and applications of Trichoderma as a biocontrol microbe. Chinese Journal of Biological Control, 2011, 27(2): 145c151.
[7] 张婷,衣思瑶,武向文,等.农药与木霉菌粉剂复配包衣种子对玉米出苗与生长的影响.上海交通大学学报(农业科学版), 2011, 29(4): 61-66. Zhang T, Yi S Y, Wu X W, et al. Effect of seed coated with complex formulation of pesticides and Trichoderma biomass on maize emergence and seedling growth. Journal of Shanghai Jiaotong University(Agricultural Science), 2011, 29(4): 61-66.
[8] Lu C G, Liu W C, Qiu J Y, et al. Identification of an antifungal metabolite produced by a potential biocontrol actinomyces strain A01. Braz J Microbiol, 2008 (39): 701-707.
[9] 梁景乐,吕忠良,赵爱华,等.双波长紫外分光光度法快速测定发酵液中纳他霉素含量.食品与发酵工业, 2007, 33(4): 127. Liang J L, Lv Z L,Zhao A H, et al. Rapid determination of natamycin concentration in the fermentation broth with dual-wavelength UV spectrophotometry. Food and Fermentation Industries, 2007, 33(4): 127.
[10] Narayana K J, Muvra V. Chitinase production by Streptomyces SP. ANU 6277. Brazilian Journal of Microbiology, (2009) 40: 725-733.
[11] 程良英.纳他霉素高产菌株的构建及其发酵工艺优化.杭州: 浙江大学, 2010. Cheng Y L. Construction of the high natamycin-producing strains and optimization on their fermentation conditions. Hangzhou: Zhejiang University, 2010.
[12] 范玲.微生物农药研究进展及产业发展对策.中国生物工程杂志,2002, 22(5):83-86. Fan L. Progress in microbial insecticides and countermeasure on industrial devolopment. China Biotechnology, 2002, 22(5):83-86. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
