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| Study on Measurement Method of Rhodobacter Sphaeroides Based on Plate Counting |
| LI Long-quan1,2,WANG Zi-quan2,LIU Si-yuan2,ZHOU Guo-ping1,SUI Zhi-wei2,ZHANG Ling2,ZHEN Xiao-xiao3,HUANG Wen-feng2,XU Qian2,4 |
1. School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, Hubei 430023, China
2. Center for Advanced Measurement Science, National Institute of Metrology, Beijing 100029, China
3. Beijing Institute of Metrology, Beijing 100029, China
4. College of Life Sciences,Hunan Normal University, Changsha, Hunan 410006, China |
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Abstract Rhodobacter sphaeroides is one of the main microorganisms producing coenzyme Q10. The quantitation detection is of great significance for optimizing the fermentation conditions of Rhodobacter sphaeroides. In this study, the plate counting method of Rhodobacter sphaeroides was optimized by comparing the inoculation method, the inoculation volume, the type of culture medium, and the culture temperature. Additionally, the optimized method was validated by collaborative experiments and uncertainty was analyzed. The final optimization result of the plate counting method is: the inoculation method is coating method, the inoculation amount is 50μL, the culture medium is Nutrient Agar, and the culture temperature is 32℃. The optimized method possessed a good repeatability and reproducibility, of which the RSD of within-laboratory repeatability is 6.0%, and inter-laboratory RSD is 16.4%. With relative expanded uncertainty of 9.2% (k=2), the optimized method could play an important role in the quantitation of Rhodobacter sphaeroides.
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Received: 18 February 2021
Published: 08 March 2023
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|
|
|
| ［9］ |
韩庆莉, 赵志瑞, 周永双, 等. 甘薯淀粉废水培养类球红细菌条件优化［J］. 安徽农学通报, 2012, 18(13):35-38.
|
| ［10］ |
周勇, 郑毅. 类球红细菌辅酶Q10发酵培养基的优化［J］. 海峡科学, 2016(3):9-11.
|
|
Zhou Y, Zheng Y. Optimization of fermentation medium for coenzyme Q10 of Rhodobacter sphaeroides［J］. Straits Science, 2016(3):9-11.
|
| ［17］ |
ISO Guide 35:2017 Reference materials—Guidance for characterization and assessment of homogeneity and stability［S］. 2017.
|
| ［19］ |
韩永志. 统计学在理化检验中的应用 第四讲 测量方法的重复性及复现性［J］. 理化检验(化学分册), 1999(12):571-572.
|
| ［1］ |
陈芳芳. 基于专利情报分析的全球辅酶Q10技术发展状况研究［J］. 中国发明与专利, 2019, 16(2):52-58.
|
| ［8］ |
张娜, 任丹, 李玉静. 关于乳与乳制品中细菌总数的检测技术研究［J］. 现代食品, 2018(13):81-82.
|
| ［16］ |
GB/T 6379.2—2004 测量方法与结果的准确度(正确度与精密度)第2部分:确定标准测量方法重复性与再现性的基本方法［S］.
|
| ［21］ |
全浩, 韩永志. 标准物质及其应用技术［M］.2版. 北京:中国标准出版社, 2003.
|
| ［4］ |
Lu W Q, Ye L D, Lü X M, et al. Identification and elimination of metabolic bottlenecks in the quinone modification pathway for enhanced coenzyme Q10 production in Rhodobacter sphaeroides［J］. Metabolic engineering, 2015, 29:208-216
|
|
Fu J L, Qian D W, Wu C Q, et al. Screening of high CoQ10 producing Rhodobacter sphaeroides by chemical mutagenesis and its fermentation medium optimization［J］. China Brewing, 2016, 35(6):90-95.
|
|
Xu M, Jiang X Z, Huang J Z, et al. Reinforcement of Rhodobacter sphaeroides cofactor NADPH to increase the production of farnesol［J］. Chinese Journal of Biotechnology, 2020, 36(1):90-99.
|
|
Xie X, Yang J L, Liu X Y. Optimizing the Determination Conditions of Total Bacterial Colonies in Soil［J］. Chinese Agricultural Science Bulletin, 2020, 36(11):92-95.
|
|
Xue L, Lin J, Sui Z W, et al. Preparation of Reference Material for Quantitative Detection of Escherichia coli［J］. Acta Metrologica Sinica, 2015, 36(6):652-656.
|
| ［7］ |
Laverty D J, Kury A L, Kuksin D, et al. Automated quantification of budding Saccharomyces cerevisiae using a novel image cytometry method［J］. Journal of Industrial Microbiology & Biotechnology, 2013, 40(6):581-588.
|
| ［12］ |
谢夏, 杨建兰, 刘新育. 土壤中细菌菌落总数的测定条件优化［J］. 中国农学通报, 2020, 36(11):92-95.
|
| ［20］ |
Parenteral Drug Association. Technical Report No.33: evaluation, validation and implementation of new as microbiological testing methods［R］. PDA, 2000.
|
| ［6］ |
扶教龙, 钱大伟, 吴晨奇, 等. 高产辅酶Q10类球红细菌的化学诱变筛选及其发酵培养基优化［J］. 中国酿造, 2016, 35(6):90-95.
|
| ［11］ |
许曼, 江贤章, 黄建忠, 等. 强化类球红细菌辅因子NADPH再生以提高法尼醇的产量［J］. 生物工程学报, 2020, 36(1):90-99.
|
| ［18］ |
阚莹, 卢晓华, 孟凡敏, 等. JJF1343—2012《标准物质定值的通用原则及统计学原理》的使用［J］. 中国计量, 2012(11):122-124.
|
|
Chen M F, Fu B Q, Lin J, et al. Development of Quantitative Reference Materence of Enterobacter Cloacae［J］. Acta Metrologica Sinica, 2017, 38(5):527-531.
|
|
Zhang N, Ren D, Li Y J. Study on Detection Technology of Total Bacteria in Milk and Dairy Products［J］. Modern Food, 2018(13):81-82.
|
| ［22］ |
隋志伟, 刘晓夏, 王晶, 等. 粘质沙雷氏菌测量方法研究［J］. 计量学报, 2018, 39(4):588-592.
|
| ［24］ |
陈敏璠, 傅博强, 林婧, 等. 阴沟肠杆菌定量标准物质的研制［J］. 计量学报, 2017, 38(5):527-531.
|
|
Chen F F. Research on Development of Global Coenzyme Q10 Technology Based on Patent Intelligence Analysis［J］. China Invention & Patent, 2019, 16(2):52-58.
|
| ［2］ |
Dhanasekaran M, Ren J. The Emerging Role of Coenzyme Q10 in Aging, Neurodegeneration, Cardiovascular Disease, Cancer and Diabetes Mellitus［J］. Current Neurovascular Research, 2005, 2(5):447-459.
|
| ［3］ |
Littarru G P, Tiano L. Clinical aspects of coenzyme Q10: An update［J］. Current Opinion in Clinical Nutrition and Metabolic Care, 2005, 8(6):641-646.
|
| ［5］ |
Yen H W, Shih T Y. Coenzyme Q10 production by Rhodobacter sphaeroides in stirred tank and in airlift bioreactor［J］. Bioprocess and Biosystems Engineering, 2009, 32(6):711-716.
|
|
Han Q L, Zhao Z Y, Zhou Y S, et al. Optimization of Fermentation Conditions for Rhodobacter sphaeroides by Sweet Potato Starch Wastewater［J］. Anhui Agricultural Science Bulletin, 2012, 18(13):35-38.
|
| ［13］ |
Tong X M, Oh Eun K, Lee B H, et al. Production of long-chain free fatty acids from metabolically engineered Rhodobacter sphaeroides heterologously producing periplasmic phospholipase A2 in dodecane-overlaid two-phase culture［J］. Microbial Cell Factories, 2019, 18(1):20.
|
| ［23］ |
薛蕾, 林婧, 隋志伟, 等. 大肠杆菌定量检测用标准物质的研制［J］. 计量学报, 2015, 36(6):652-656.
|
| ［14］ |
Mougiakos I, Orsi E, Ghiffary M R, et al. Efficient Cas9-based genome editing of Rhodobacter sphaeroides for metabolic engineering［J］. Microbial Cell Factories, 2019, 18(4):89-561.
|
| ［15］ |
Chen Y G, Li M, Meng F S, et al. Optimal poly (3-hydroxybutyrate/3-hydroxyvalerate) biosynthesis by fermentation liquid from primary and waste activated sludge［J］. Environmental Technology, 2014, 35(13-16):1791-801.
|
|
Sui Z W,Liu X X,Wang J, et al. Study on Measurement Method of Serratia Marcescens［J］. Acta Metrologica Sinica, 2018, 39(4):588-592.
|
|
|
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2023, Vol. 44 
