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Integrated Expression of the oenococcus oeni mleA Gene in Saccharomyces cerevisiae

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Agricultural Sciences in China 2009年第7期8卷 821-827页

作者： LIU Yan-lin LI Hua College of Enology Northwest A ＆F University/Shaanxi Engineering Research Center for Viti-Viniculture Yangling 712100 P.R. China

Malolactic enzyme is the function enzyme which catalyses the reaction for L-malate converting to L-lactic during malolactic fermentation （MLF）. In this paper, researches concerning the malolactic enzyme gene mleA cloned from a patent strain oenococcus oeni SD-2a screened in Chinese wine and integrated expressing in Saccharomyces cerevisiae were performed in order to carry out both alcoholic fermentation （AF） and malolactic fermentation （MLF） during winemaking, with a view to achieving a better control of MLF in enology. To construct the expression plasmid named pYILmleA, cloned mleA gene, PGK1 promoter, and ADH1 terminator were ligated and inserted into integrating vector YIp5. Yeast transformants were screened on SD/-Ura and identified by auxotrophic test, mating type test, and colony PCR. Target protein was detected by SDS-PAGE and the targeted gene integrated to the chromosome was detected by dot bloting hybridization. After the transformant was cultured in SD/-Ura adding glucose （10%） and L-malate （5 648 mg L-1） for 4 d, the culture supernatant was collected and L-malate and L-lactic acid contents were detected by HPLC. 1 278-1 312 mg L-1 L-lactic acids were detected, while the comparative drop rates of L-malate were 20.18-20.85%. L-malate and L-lactic contents of the transformants showed extra significant difference and significant difference with the control ones by t-test respectively. The result indicated that the functional expression was achieved in recombinants S. cerevisiae.

关键词： oenococcus oeni malolactic enzyme gene integrated recombinant expression Saccharomyces cerevisiae

Advanced Progress on Adaptive Stress Response of oenococcus oeni

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Journal of Northeast Agricultural University(English Edition) 2013年第1期20卷 91-96页

作者： Zhao Wen-ying Kang Zhen-kui College of Chemical Engineering and Environment North University of China Tianjiaohong Biotechnology Company

Oenoccoccus oeni is an alcohol-tolerant, acidophilic lactic acid bacterium with its ability to perform malolactic fermentation in wine, which is of fundamental importance in oenology. As a representative of the wine bacterium with remarkable adaptability to the very harsh physicochemical conditions of wine, many studies were carded out for its applied interest and focused mainly on its stress response mechanisms of O. oeni. on both physiological and molecular levels. In this review, three main stress response mechanisms in O. oeni during culturing process were addressed. Of them, various solute transporters and secondary metabolic energy-generating systems were utilized to control the intracellular environment and the energetic status of O. oeni. The changes in cell membrane fatty acid composition profiles and synthesis of stress proteins, especially small heat shock proteins were required for active cell response to maintain membrane integrity and function under stress conditions. The study on stress response of O. oeni played an important role on culture bacteria selection, making inoculation culture and construction of other engineering bacteria.

关键词： oenococcus oeni stress response acid stress alcohol stress

Isolation and Screening of oenococcus oeni and Its Medium Optimization

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Agricultural Biotechnology 2021年第1期10卷 114-115,119页

作者： Dongdong HOU Xi HE College of Biological Engineering Qilu University of Technology(Shandong Academy of Sciences)Jinan 250353China

[Objectives] This study was conducted to further improve the application of oenococcus oeni in the wine brewing industry. [Methods] O. oeni was isolated, screened, and identified, and medium optimization was performed to determine the best medium, temperature and oxygen conditions. [Results] O. oeni grew better in mFT80 medium, and the culture conditions were 25 ℃ and an anaerobic condition. [Conclusions] This study provides a reference for the development of China’s wine industry.

关键词： oenococcus oeni Isolation and screening Culture medium Optimization

Microbiota for production of wine with enhanced functional components

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Food Science and Human Wellness 2023年第5期12卷 1481-1492页

作者： Armachius James Ting Yao Hengming Ke Yousheng Wang Beijing Advanced Innovation Centre for Food Nutrition and Human Health Beijing Technology and Business UniversityBeijing 100048China Tanzania Agricultural Research Institute Makutupora Centre P.O.Box 1676DodomaTanzania Rizhao HUAWEI Institute of Comprehensive Health Industries Shandong KEEPFIT Biotech.Co.Ltd.Rizhao 276800China Department of Biochemistry and Biophysics and Lineberger Comprehensive Centre The University of North CarolinaChapel Hill 27599USA

Microbial communities during winemaking are diverse and change throughout the fermentation process. Microorganisms not only drive alcohol fermentation, flavor and aroma, but also enhance wine functional components such as extraction of polyphenols from the berries, production of γ-aminobutyric acid, hydroxytyrosol and melatonin. Polyphenols such as resveratrol, catechin and quercetin determine the functional quality of the wine. Moderate wine consumption, particularly red wine has been associated with functional benefits to human health, which includes anti-inflammation, promoting healthy aging, prevention of cardiovascular diseases, cancers, type 2 diabetes and metabolic syndrome. Indeed, the management of microbiota allows the production of wine with distinct features and functional components that benefits human health. This review scrutinizes the possible contributions of wine microbiota to the production of wine with enhanced functional components and highlights the contributions of Saccharomyces and non-Saccharomyces yeasts and bacteria to enhance wine functional components during winemaking. Thus, contributing to the dissemination of the benefits of light to moderate wine intake to human health.

关键词： Wine Polyphenols Resveratrol oenococcus oeni Non-Saccharomyces yeast