Expression of food-grade phytase in Lactococcus lactis from optimized conditions in milk broth(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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概要

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• The major objective of this study was to engineer lactic acid bacteria to produce the enzyme phytase from a gene native to Bacillus subtilis GYPB04. The phytase gene (phyC) of B. subtilis GYPB04 was cloned into the plasmid pMG36e for expression in Lactococcus lactis. The enzyme activity in L lactis cultured in GM17 broth was 20.25 U/mL at 36℃. The expressed phytase was characterized as active in a pH range of 2.0-9.0 at a temperature range of 20-80℃, with an optimum pH of 5.5-6.5 and temperature of 60℃. When cultured in food-grade milk broth, the transformed L lactis grew to an OD_<600> nm value of 1.05 and had a phytase yield of 13.58 U/mL. In same broth under optimized conditions for cell growth and phytase production, the transformant reached an OD_<600> nm value of 1.68 and a phytase yield of 42.12 U/mL, representing approximately 1.6-fold and 3.1-fold increases, respectively, compared to growth in natural milk broth. Fermentation was scaled to 5 L under optimized conditions, and product analysis revealed a final OD_<600> nm value of 1.89 and an extracellular enzyme activity of 24.23 U/mL. The results of this study may be used in the dairy fermentation industry for the development of functional, healthy yogurts and other fermented dairy foods that provide both active phytase and viable probiotics to the consumer.

• 2013-07-00

著者

• CAO Yi

  Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Science,

• Xu Hui

  Microbiology And Metabolic Engineering Key Laboratory Of Sichuan Province College Of Life Science Si

• Fei Baojin

  Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Science, Sichuan University

• Qiao Dairong

  Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Science, Sichuan University

• Fei Baojin

  Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University

• Miao Yuzhi

  College of Life Sciences, Sichuan Normal University

• Miao Yuzhi

  College of Life Sciences, Sichuan Normal University:Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University

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