Fermentation experiments to produce validamycins from crude substrates by Streptomyces hygroscopicus were carried out in an external-loop airlift bioreactor (0.0115 m^3 ) with a low ratio of height to diameter of the ...Fermentation experiments to produce validamycins from crude substrates by Streptomyces hygroscopicus were carried out in an external-loop airlift bioreactor (0.0115 m^3 ) with a low ratio of height to diameter of the riser of 2.9 and a ratio of riser to downcomer diameter of 6.6. The influences of gas flow rate and liquid volume on fermentation of validamycins were investigated. Comparisons of validamycin fermentation were made among the external-loop airlift bioreactor, a mechanically stirred tank bioreactor (0.010m^3 ) and shaking flasks. Under the same operation conditions including fermentation medium composition, inoculum ratio and culture temperature, the fermentation time in the external-loop airlift bioreactor (45 h) was shorter than that in the shaking flasks (100 h) and the same as that in the mechanically stirred tank bioreactor. After a total fermentation time of 45 h under optimized operation conditions, average validamycin concentration obtained in the external-loop airlift bioreactor was close to 19630 μg·ml^-1 validamycin concentration in the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the mechanically stirred tank bioreactor in production of validamycins from crude substrates with dregs by Streptomyces hygroscopicus.展开更多
As an important medicinal intermediate with broad uses, validamine, an aminocyclitol, isolated from the enzymolysis broth of validamycins, has gained more and more attention. The absolute configuration of validamine i...As an important medicinal intermediate with broad uses, validamine, an aminocyclitol, isolated from the enzymolysis broth of validamycins, has gained more and more attention. The absolute configuration of validamine is similar to that of a-D-glucose, and it demonstrates powerful inhibition activity on glycosidase. In this paper, the inhibitory effect of validamine on porcine small intestinal sucrase was investigated. Validamine was found to be a potent, competitive inhibitor to porcine small intestinal sucrase in vitro with an IC50 value of 6.85 X 10(-4) mol center dot L-1. Validamine exhibited a dose-dependent inhibition effect on porcine small intestinal sucrase, whereby the inhibition interaction of validamine and porcine small intestinal sucrase was a fast binding process. The inhibition of validamine on porcine small intestinal sucrase was pH-dependent.展开更多
Cultivation shift from 30℃ to 37℃ significantly enhanced validamycin (VAL) production. Analyzed by reverse-transcription PCR, the transcription of three val genes, valA, valK and valG, representing the three operons...Cultivation shift from 30℃ to 37℃ significantly enhanced validamycin (VAL) production. Analyzed by reverse-transcription PCR, the transcription of three val genes, valA, valK and valG, representing the three operons of the cluster was simultaneously increased at elevated temperature. Furthermore, the transcription of valP and valQ, a pair of two-component regulators in validamycin biosynthetic gene cluster, was also increased at 37℃. Inactivation of valP and valQ reduced validamycin production at 37℃ to the yield level of wild type strain at 30℃, and the val genes showed reduced expression in the mutant LL-8 at 37℃. These results revealed that the two-component regulator valP and valQ contribute to the elevated validamycin production.展开更多
基金Supported by the Natural Science Foundation of Zhejiang Province (No. ZB0106)the National Natural Science Foundation of China (No. 20176055).
文摘Fermentation experiments to produce validamycins from crude substrates by Streptomyces hygroscopicus were carried out in an external-loop airlift bioreactor (0.0115 m^3 ) with a low ratio of height to diameter of the riser of 2.9 and a ratio of riser to downcomer diameter of 6.6. The influences of gas flow rate and liquid volume on fermentation of validamycins were investigated. Comparisons of validamycin fermentation were made among the external-loop airlift bioreactor, a mechanically stirred tank bioreactor (0.010m^3 ) and shaking flasks. Under the same operation conditions including fermentation medium composition, inoculum ratio and culture temperature, the fermentation time in the external-loop airlift bioreactor (45 h) was shorter than that in the shaking flasks (100 h) and the same as that in the mechanically stirred tank bioreactor. After a total fermentation time of 45 h under optimized operation conditions, average validamycin concentration obtained in the external-loop airlift bioreactor was close to 19630 μg·ml^-1 validamycin concentration in the mechanically stirred tank bioreactor. It was demonstrated that the external-loop airlift bioreactor could substitute for the mechanically stirred tank bioreactor in production of validamycins from crude substrates with dregs by Streptomyces hygroscopicus.
基金Supported by the National Fund of the Major Basic Research Development Program (973 Project) (No.2003CB716005)the National Natural Science Foundation of China (No. 20176055) the Natural Science Foundation of Zhejiang Province (No. ZB0106).
文摘As an important medicinal intermediate with broad uses, validamine, an aminocyclitol, isolated from the enzymolysis broth of validamycins, has gained more and more attention. The absolute configuration of validamine is similar to that of a-D-glucose, and it demonstrates powerful inhibition activity on glycosidase. In this paper, the inhibitory effect of validamine on porcine small intestinal sucrase was investigated. Validamine was found to be a potent, competitive inhibitor to porcine small intestinal sucrase in vitro with an IC50 value of 6.85 X 10(-4) mol center dot L-1. Validamine exhibited a dose-dependent inhibition effect on porcine small intestinal sucrase, whereby the inhibition interaction of validamine and porcine small intestinal sucrase was a fast binding process. The inhibition of validamine on porcine small intestinal sucrase was pH-dependent.
基金Supported by National Key Basic Research and Development Program of China (Grant No. 2003CB114205)National High Technology Research and Development Program of China (Grant No. 2006AA02Z185)+1 种基金the National Natural Science Foundation of China (Grant No. 30570019)the Shanghai Municipal Council of Science and Technology, and Shanghai Leading Academic Discipline Project B203
文摘Cultivation shift from 30℃ to 37℃ significantly enhanced validamycin (VAL) production. Analyzed by reverse-transcription PCR, the transcription of three val genes, valA, valK and valG, representing the three operons of the cluster was simultaneously increased at elevated temperature. Furthermore, the transcription of valP and valQ, a pair of two-component regulators in validamycin biosynthetic gene cluster, was also increased at 37℃. Inactivation of valP and valQ reduced validamycin production at 37℃ to the yield level of wild type strain at 30℃, and the val genes showed reduced expression in the mutant LL-8 at 37℃. These results revealed that the two-component regulator valP and valQ contribute to the elevated validamycin production.
