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食品与发酵工业  2022, Vol. 48 Issue (3): 9-14    DOI: 10.13995/j.cnki.11-1802/ts.028337
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
毕赤酵母产类人胶原蛋白发酵条件的优化
冯梦茹, 周化岚, 张建国*
(上海理工大学 医疗器械与食品学院,食品科学与工程研究所,上海,200093)
Optimization of human-like collagen expression by recombinant Komagataella phaffii
FENG Mengru, ZHOU Hualan, ZHANG Jianguo*
(Institute of Food Science and Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China)
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摘要 胶原蛋白是结缔组织中一种重要蛋白质,具有抗皱、抗老化和补钙壮骨的功效,在食品工业中应用广泛。由于胶原蛋白的需求量大,因此利用重组微生物生产胶原蛋白成为近年来的研究热点。该文采用重组有类人胶原蛋白基因的毕赤酵母,通过优化培养基成分,使其在无机盐培养基中表达胶原蛋白。首先,类人胶原蛋白在每24 h补加体积分数1.5%甲醇诱导120 h后的产量可达18.87 mg/L,进而将无机盐培养基中盐浓度降至1/3,有效避免了类人胶原蛋白的水解,也减少了重组毕赤酵母菌株分泌的杂蛋白量。进一步添加1 g/L L-脯氨酸等辅助营养物使类人胶原蛋白的产量提高至40.26 mg/L,为初始类人胶原蛋白表达量的2倍。结果表明,添加氨基酸等氮源能显著提高重组毕赤酵母的类人胶原蛋白分泌量,为重组类人胶原蛋白的发酵生产提供了指导。
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冯梦茹
周化岚
张建国
关键词:  毕赤酵母  甲醇诱导  类人胶原蛋白  辅助氮源  发酵条件优化    
Abstract: Collagen is an important structural protein in connective tissue, plays an important role in anti-wrinkle, anti-aging and calcium reinforcement, and widely applies in food industry. This research optimized the cultivation condition of recombinant Komagataella phaffii to express human-like collagen in an inorganic salt medium. Firstly, the yield of human-like collagen was 18.87 mg/L at 1.5% methanol induction per 24 h after 120 h induced fermentation. Further, degradation of human-like collagen was avoided by reducing the salts concentration to 1/3 of initial value, also leading to less impure protein in fermentation broth. The yield of human-like collagen reached 40.26 mg/L at the condition 1 g/L L-proline addition, which was 2-fold of initial yield of human-like collagen. This research indicated that the nitrogen source, such as amino acid, increased the yield of human-like collagen by recombinant K. phaffii significantly, which provides an important direction for collagen production through fermentation.
Key words:  Komagataella phaffii    methanol induction    human-like collagen    supplemental nitrogen resource    fermentation optimization
收稿日期:  2021-07-13      修回日期:  2021-07-30           出版日期:  2022-02-15      发布日期:  2022-03-04      期的出版日期:  2022-02-15
基金资助: 国家自然科学基金项目(31870045)
作者简介:  硕士研究生(张建国教授为通信作者,E-mail:jgzhang@usst.edu.cn)
引用本文:    
冯梦茹,周化岚,张建国. 毕赤酵母产类人胶原蛋白发酵条件的优化[J]. 食品与发酵工业, 2022, 48(3): 9-14.
FENG Mengru,ZHOU Hualan,ZHANG Jianguo. Optimization of human-like collagen expression by recombinant Komagataella phaffii[J]. Food and Fermentation Industries, 2022, 48(3): 9-14.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.028337  或          http://sf1970.cnif.cn/CN/Y2022/V48/I3/9
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