作 者：
姜艳平;吴煜;张希;姜新鹏;唐丽杰;乔薪瑗;崔文;李一经
关键词：
犬γ干扰素;发酵罐;条件优化;乳酸乳球菌
摘 要：
为探讨发酵罐培养重组乳酸乳球菌表达犬γ-干扰素最佳条件,研究测定重组乳酸乳球菌p AMJ399-Ca IFN-γ/PSM565静置培养时生长曲线及p H,与添加不同浓度β甘油磷酸二钠的生长曲线及p H进行比较。结果表明,β-甘油磷酸二钠作为缓冲溶液,能使培养细菌p H保持稳定,促进重组菌生长。对重组乳酸乳球菌p AMJ399-Ca IFN-γ/PSM565在发酵罐中的培养条件优化,并对不同p H条件下培养重组乳酸乳球菌OD600及重组p AMJ399-Ca IFN-γ/PSM565表达的蛋白量进行Western-blot检测。发酵结果显示,32℃培养,搅拌子转速为52r·min-1,6 h后以0.25 m L·min-1速率补充50%葡萄糖,明显提高菌群数量,p H在5.5和5.75时检测重组菌OD600值达到6.87和7.0。p H为5.5和5.75时培养诱导重组乳酸乳球菌p AMJ399-Ca IFN-γ/PSM565的蛋白表达量均比p H 6和p H 6.25时高,但与静止培养诱导结果相比,重组菌表达量增长不明显。研究可为工业化生产重组犬γ-干扰素提供技术支持。
译 名：
Optimization of the fermentation condition of expressing the γ-interferon in recombinant Lactococcus lactis
作 者：
JIANG Yanping;WU Yu;ZHANG Xi;JIANG Xinpeng;TANG Lijie;QIAO Xinyuan;CUI Wen;LI Yijing;School of Veterinary Medicine,Northeast Agricultural University;
关键词：
γ-interferon of canine;;fermenter;;optimization;;Lactococcus lactis
摘 要：
In order to explore the best conditions in the expressing canine γ- interferon in recombinant Lactococcus lactis. This study determined the static culture of p H value and the growth curve in the recombinant Lactococcus lactis of p AMJ399-Ca IFN-γ/PSM565, comparing the deferent concentration of β- glycerophosphate disodium and p H curve. The results showed that the β- glycerophosphate disodium, the culture of recombinant Lactococcus lactis, could play the role of buffer, which can keep the stabilization of p H. The culture conditions of recombinant Lactococcus lactis of p AMJ399-Ca IFN-γ/PSM565 were optimized in the fermentation tank. The expression of Ca IFN-γ in recombinant Lactococcus lactis of p AMJ399- Ca IFN- γ/PSM565 was tested in different OD600 and p H with Westernblot. Fermentation results showed the induction at after 6 h, could increase the cultures of colony forming units. The OD600 of recombinant 37 ℃, incubating with shaking at 52 r· min- 1, and 50% glucose was added to a speed of 0.25 m L· min-1Lactococcus lactis could get from 6.87 to 7.0 in p H between 5.5and 5.75, which was higher for the expression of p AMJ399- Ca IFN- γ/PSM565 than the p H 6 and p H6.25. But there were no significant differences of inducing the expression protein comparing with static culture. This study increased the expression of p AMJ399- Ca IFN- γ/PSM565, building the technical assurance for the industrialization, and establishing the material foundation.
