| 评价芽孢杆菌脂肽产量及底物碳利用效率的新方法 |
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| 引用本文： | 王晶,刘茗,冯歌林,方伟,高竞,梁辰飞,秦华,陈俊辉,徐秋芳.评价芽孢杆菌脂肽产量及底物碳利用效率的新方法[J].浙江农林大学学报,2019,36(6):1182-1189. |
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| 作者姓名： | 王晶 刘茗 冯歌林 方伟 高竞 梁辰飞 秦华 陈俊辉 徐秋芳 |
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| 作者单位： | 1.浙江农林大学 浙江省森林生态系统碳循环与固碳减排重点实验室, 浙江 杭州 3113002.浙江农林大学 环境与资源学院, 浙江 杭州 311300 |
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| 基金项目： | 浙江省省院合作林业科技项目2017SY01浙江省科学技术重点研发计划项目2015C02G4010091 |
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| 摘 要： | 芽孢杆菌Bacillus代谢产生的脂肽类物质具有抗菌、抗肿瘤、抗病毒等多种生物活性，但芽孢杆菌脂肽产量普遍较低。探索高产量的培养条件是开发利用的重点，而简单快速的产量评价方法则可大大提高条件探索的效率。以解淀粉芽孢杆菌Bacillus amyloliquefaciens WK1为试验菌株，首先通过含发酵液培养基对病原菌的抑制实验确定最佳培养时间，然后设计初始pH值分别为5，6和7的培养基生产脂肽物质，通过测定发酵液酸沉降前后总有机碳的差值，推算脂肽类物质粗提物的产量（差减法），以传统的重量法为对照。结果表明:试验所用的解淀粉芽孢杆菌WK1菌株的最佳发酵时间为72 h。重量法与差减法所测得的不同处理脂肽物质产量差异规律相同，从高到低依次为pH 6，pH 7，pH 5，两者的相关性系数为0.563 2（n=9），但相同初始pH值条件下差减法所得的脂肽物质产量均高于重量法。初始pH值为5条件下脂肽物质合成效率和二氧化碳排放量均最低，初始pH值为6与pH值为7条件下二氧化碳排放量相同，为700 mg·kg-1，但初始pH值为6条件下产脂肽物质产量（1 228 mg·L-1）和脂肽物质合成效率（234 mg·kg-1）最高。可见，差减法是评价脂肽产量的简单快速而有效的方法。
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| 关 键 词： | 微生物学 解淀粉芽孢杆菌 脂肽类产量 差减法 重量法 底物碳利用 |
| 收稿时间： | 2018-11-14 |
A new evaluation method of lipopeptide production and substrate carbon utilization efficiency using the WK1 strain of Bacillus amyloliquefaciens |
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| WANG Jing,LIU Ming,FENG Gelin,FANG Wei,GAO Jing,LIANG Chenfei,QIN Hua,CHEN Junhui,XU Qiufang.A new evaluation method of lipopeptide production and substrate carbon utilization efficiency using the WK1 strain of Bacillus amyloliquefaciens[J].Journal of Zhejiang A&F University,2019,36(6):1182-1189. |
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| Authors: | WANG Jing LIU Ming FENG Gelin FANG Wei GAO Jing LIANG Chenfei QIN Hua CHEN Junhui XU Qiufang |
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| Affiliation: | 1.Zhejiang Provincial Key Laboratory of Carbon Cycling in Forest Ecosystems and Carbon Sequestration, Zhejiang A & F University, Hangzhou 311300, Zhejiang, China2.College of Environmental and Resource Sciences, Zhejiang A & F University, Hangzhou 311300, Zhejiang, China |
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| Abstract: | The lipopeptide produced by Bacillus amyloliquefaciens has a variety of biological activities such as antibacterial, anti-tumor, and anti-virus; however, its production is generally very low. To explore better fermentation conditions so as to gain more lipopeptide with easy and fast evaluation methods for lipopeptide production that would greatly improve work efficiency, the yield of crude lipopeptide based on the difference of total organic carbon (TOC) in a liquid before and after lipopeptide separation was calculated using the acid deposition approach. First, optimal fermentation time for producing lipopeptide by pathogen inhibiting culture on plates containing a fermentation liquid was determined. Then, a liquid medium with pH 5, 6, and 7, for producing different concentrations of lipopeptide, was prepared. Also, CO2 emission rates for microbial activity in the pH 6 and 7 treatments were measured. Results showed that 72 hours was the optimal fermentation time. The crude yield of lipopeptide calculated from the difference of TOC in liquid before and after lipopeptide separation was positive compared to traditional weight methods with the coefficient being 0.563 2 (n=9). For both methods, pH 6 > pH 7 > pH 5. However, for the same pH of the liquid medium, crude yield was lower with weight methods than the TOC difference methods. Lipopeptide production efficiency and CO2 emissions rate with the WK1 strain were lowest for the pH 5 treatment. However, crude yield of lipopeptide (in a 1 228 mg·L-1 medium) and lipopeptide production efficiency (in a 234 mg·kg-1 OC medium) were highest at pH 6. Thus, the TOC difference method was an easy and fast method for lipopeptide production evaluation. |
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