Abstract: To improve production efficiency of 2-phenylethanol (2PE) from L-Pheby Metschnikowia sp., a high-yield 2PE strain CICC 1467 was screened from four candidate Metschnikowia strains. Then, factors were investigated to enhance the production of 2PE, including L-Phe dosage, carbon source, initial pH, temperature, and inoculation volume. Furthermore, the fermentation ability of CICC 1467 under non-sterile conditions was investigated. Under optimal conditions, the final 2PE and molar yield reached 2.24 g/L and 0.76, respectively. The condition were as follows: L-Phe 4 g/L, glucose 50 g/L, initial pH 3.0, 25℃, inoculation volume 5%, 44 h, and with non-sterilized tap water as the solvent. The results represent the highest molar yield compared with the ever reported of 2PE from L-Phe by Metschnikowia. The study proved that Metschnikowia sp. had the ability to synthesize 2PE efficiently, and important reference value for microbial synthesis of 2PE.
HUA D L,XU P.Recent advances in biotechnological production of 2-phenylethanol[J].Biotechnology Advances,2011,29(6):654-660.
[2]
国家卫生和计划生育委员会.GB 2760—2014食品添加剂使用标准[S].北京:中国标准出版社,2014.National Health and Family Planning Commission.GB 2760—2014 National food safety standard for uses of food additive[S].Beijing:Standards Press of China,2014.
[3]
NEGRE F,KISH C M,BOATRIGHT J,et al.Regulation of methylbenzoate emission after pollination in snapdragon and Petunia flowers[J].Plant Cell,2003,15(12):2 992-3 006.
[4]
TIEMAN D,TAYLOR M,SCHAUER N,et al.Tomato aromatic amino acid decarboxylases participate in synthesis of the flavor volatiles 2-phenylethanol and 2-phenylacetaldehyde[J].Proceedings of the National Academy of Sciences,2006,103(21):8 287-8 292.
[5]
戴琳, 单银花,王志祥,等.超临界CO2萃取玫瑰精油的工艺优化[J].食品工业科技,2015,36(3):266-269;274.DAI L,SHAN Y H,WANG Z X,et al.Technical optimization on supercritical CO2 extraction of rose essential oil[J].Science and Technology of Food Industry,2015,36(3):266-269;274.
[6]
YAMAGUCHI K,EBITANI K,KANEDA K.Hydrotalcite-catalyzed epoxidation of olefins using hydrogen peroxide and amide compounds[J].The Journal of Organic Chemistry,1999,64(8):2 966-2 968.
[7]
陈先锐, 王肇悦,何秀萍.酵母菌合成2-苯乙醇的研究进展[J].生物工程学报,2016,32(9):1 151-1 163.CHEN X R,WANG Z Y,HE X P.Advances in biosynthesis of 2-phenylethanol by yeasts[J].Chinese Journal of Biotechnology,2016,32(9):1 151-1 163.
[8]
QIAN X J,YAN W,ZHANG W M,et al.Current status and perspectives of 2-phenylethanol production through biological processes[J].Critical Reviews in Biotechnology,2019,39(2):235-248.
[9]
WANG Y Q,ZHANG Z Y,LU X Y,et al.Transcription factor Hap5 induces gsh2 expression to enhance 2-phenylethanol tolerance and production in an industrial yeast Candida glycerinogenes[J].Applied Microbiology and Biotechnology,2020,104(9):4 093-4 107.
[10]
CHREPTOWICZ K,MIERZEJEWSKA J.Enhanced bioproduction of 2-phenylethanol in a biphasic system with rapeseed oil[J].New Biotechnology,2018,42:56-61.
[11]
郭孝敬, 郑晓吉,史学伟,等.高产2-苯乙醇酵母的筛选及其发酵条件优化[J].中国调味品,2019,44(11):55-61.GUO X J,ZHENG X J,SHI X W,et al.Screening of high-yield 2-phenylethanol yeast and optimization of its fermentation conditions[J].China Condiment,2019,44(11):55-61.
[12]
YAN W,ZHANG X Y,QIAN X J,et al.Comprehensive investigations of 2-phenylethanol production by high 2-phenylethanol tolerating Meyerozyma sp.strain YLG18[J].Enzyme and Microbial Technology,2020,140.DOI:10.1016/j.enzmictec.2020.109629.
[13]
黄秋容, 粟桂娇,赖芳,等.肉桂内生菌Sphingomonas sp.Z45生物转化肉桂醇生成天然2-苯乙醇[J].精细化工,2019,36(12):2 425-2 430.HUANG Q R,SU G J,LAI F,et al.Biotransformation of cinnamon alcohol into natural 2-phenylethanol by endophytic bacteria Sphingomonas sp.Z45 from cinnamon[J].Fine Chemicals,2019,36(12):2 425-2 430.
[14]
LIMA L A D,VENTORIM R Z,BIANCHINI I D A,et al.Obtainment,selection and characterization of a mutant strain of Kluyveromyces marxianus that displays improved production of 2-phenylethanol and enhanced DAHP synthase activity[J].Journal of Applied Microbiology,2020.DOI:10.1111/jam.14793.
[15]
RUIZ J,BELDA I,BEISERT B,et al.Analytical impact of Metschnikowia pulcherrima in the volatile profile of Verdejo white wines[J].Applied Microbiology and Biotechnology,2018,102(19):8 501-8 509.
[16]
CHANTASUBAN T,SANTOMAURO F,GORE-LLOYD D,et al.Elevated production of the aromatic fragrance molecule,2-phenylethanol,using Metschnikowia pulcherrima through both de novo and ex novo conversion in batch and continuous modes[J].Journal of Chemical Technology and Biotechnology,2018,93(8):2 118-2 130.
[17]
GORMMLOYD D,SUMANN I,BRACHMANN A O.Snf2 controls pulcherriminic acid biosynthesis and antifungal activity of the biocontrol yeast Metschnikowia pulcherrima[J].Molecular Microbiology,2019,112 (1):317-332.
[18]
田亚琴, 葛念念,周易,等.美极梅奇酵母抑制芒果炭疽菌的拮抗机理初探[J].食品工业科技,2018,39(1):82-86.TIAN Y Q,GE N N,ZHOU Y,et al.Primary study on antagonism mechanism of Metschnikowia pulcherrima yeast inhibiting Colletotrichum gloeosporioides in mango fruit[J].Science and Technology of Food Industry,2018,39(1):82-86.
[19]
SANTAMAURO F,WHIFFIN F M,SCOTT R J,et al.Low-cost lipid production by an oleaginous yeast cultured in non-sterile conditions using model waste resources[J].Biotechnology for Biofuels,2014,7(1):34-44.
[20]
CLEMENTE-JIMENEZ J M,MINGORANCE-CAZORLA L,MARTINEZ-RODRIGUEZ S,et al.Molecular characterization and oenological properties of wine yeasts isolated during spontaneous fermentation of six varieties of grape must[J].Food Microbiology,2004,21:149-155.
[21]
CHREPTOWICZ K,STERNICKA M K,KOWALSKA P D,et al.Screening of yeasts for the production of 2-phenylethanol (rose aroma) in organic waste-based media[J].Letters in Applied Microbiology,2018,66 (2):153-160.
[22]
LI N,LIU Y,ZHAO Y,et al.Simultaneous HPLC determination of amino acids in tea infusion coupled to pre-column derivatization with 2,4-dinitrofluorobenzene[J].Food Analytical Methods,2015,9 (5):1 307-1 314.
[23]
GATTI R.Simultaneous determination of taurine,N-acetylcysteine,glycine and methionine in commercial ormulations by high-performance liquid chromatography[J].Chromatographia,2019,82 (12):1 833-1 837.
[24]
杜闪, 王雪花,杨政茂,等.生物转化合成β-苯乙醇代谢途径及其调控的研究[J].食品与发酵工业,2014,40(1):168-173.DU S,WANG X H,YANG Z M,et al.Study on metabolic pathways of β-phenylethanol bioconversion and regulation[J].Food and Fermentation Industries,2014,40(1):168-173.
[25]
牛明福, 李亚恒,陈金帅,等.马克斯克鲁维酵母生物转化2-苯乙醇工艺优化及耐高温特性分析[J].食品与发酵工业,2018,44(2):15-20.NIU M F,LI Y H,CHEN J S,et al.Optimization and characterization of 2-phenylethanol bioconversion by thermo tolerant yeast Kluyveromyces marxianus[J].Food and Fermentation Industries,2018,44(2):15-20.
[26]
MARIS A J A,et al.The Ehrlich pathway for fusel alcohol production:a century of research on Saccharomyces cerevisiae metabolism[J].Applied and Environmental Microbiology,2008,74(8):2 259-2 266.
[27]
黄筱萍, 黄国昌,金丹凤,等.酿酒酵母静息细胞转化合成2-苯乙醇[J].食品与生物技术学报,2017,36 (12):1 316-1 323.HUANG X P,HUANG G C,JIN D F,et al.Study on bioconversion of 2-phenylethanol by Saccharomyces cerevisiae resting cell[J].Journal of Food Science and Biotechnology,2017,36 (12):1 316-1 323.
[28]
王镜岩, 朱圣庚,徐长法.生物化学教程[M].北京:高等教育出版社,2008.WANG J Y,ZHU S G,XU C F.Essential Biochemistry[M].Beijing:Higher Education Press,2008.
[29]
陈燕飞. pH对微生物的影响[J].太原师范学院学报(自然科学版),2009,8(3):121-124,131.CHEN Y F.pH to uygur biology influence[J].Journal of Taiyuan Normal University(Natural Science Edition),2009,8(3):121-124,131.
[30]
CHAROENCHAI C,FLEET G H,HENSCHKE P A.Effects of temperature,pH,and sugar concentration on the growth rates and cell biomass of wine yeasts[J].American Journal of Enology and Viticulture,1998,49(3):283-288.
[31]
贾薇, 白岩岩,冯杰,等.樟芝发酵产2-苯乙醇的条件优化[J].食用菌学报,2016,23(1):47-52.JIA W,BAI Y Y,FENG J,et al.Screening and fermentation conditions optimization of 2-phenethyl alcohol marine yeast[J].Acta Edulis Fungi,2016,23(1):47-52.