Novel spectrophotometric approach for determination of validamycin A in fermentation of Streptomyces hygroscopicus.
Validamycin A (Val-A), produced by Streptomyces hygroscopicus 5008 in industrial fermentation, is one of the most widely used anti-fungal agro-antibiotics in Asia and high performance liquid chromatography (HPLC) assay is usually used to determine the production of Val-A. A new approach to determine Val-A by spectrophotometer is developed. During the fermentation of S. hygroscopicus 5008, a pigment secretion was found along with the Val-A biosynthesis. There was a stable relationship between the concentration of Val-A and spectral absorption (SA) value of this pigment at 450 nm, even in different fermentation cultures or conditions. Using SA value as interior label, a rapid spectrophotometric method for determining Val-A production was established. In comparing Val-A productivity by HPLC method with that by SA method, the relative standard deviation (R.S.D.) was 0.007 (less than 0.05, no variation) and the conditional probability [Pr(T < t)] was 0.3491 (greater than 0.05, no difference) at the optimal time point of Val-A fermentation, which demonstrated SA method was as stable and accurate as standard HPLC method. It was applied successfully to finding positive strains with high Val-A productivity and short fermentation time. SA assay is an accurate and cost-effective method for measuring Val-A and screening high-producing strains, and this work provides a new insight for rapid quantitative analysis of antibiotics in fermentation of pigment-producing strains.
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address: jiangnanliwei2008@163.com.
Wei
W
Li
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address: fjs061@163.com.
Jinsong
J
Feng
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address: wsliuyan@163.com.
Yan
Y
Liu
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address: 2234294785@qq.com.
Jing
J
Jiang
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address: xdzheng@zju.edu.cn.
Xiaodong
X
Zheng
College of Biosystems Engineering and Food Science, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou 310058, Zhejiang, China. Electronic address: vivianzhou11@zju.edu.cn.
Wen-Wen
WW
Zhou
D000900:Anti-Bacterial Agents / Q000032:analysis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D005285:Fermentation; D007294:Inositol / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D057230:Limit of Detection; D015203:Reproducibility of Results; D013053:Spectrophotometry / Q000379:methods; D013302:Streptomyces / Q000737:chemistry* / Q000378:metabolism*
D000900:Anti-Bacterial Agents; C030790:validamycin A; D007294:Inositol
High performance liquid chromatography analysis; Pigment; Spectrophotometry; Streptomyces; Validamycin A