In order to screen non-Saccharomyces yeasts autochthonous strains endowed with high glycosidase activity, 912 yeasts were isolated from spontaneous alcoholic fermentations of the major wine grape cultivars planted in Hexi Corridor of Gansu Province. The β-glucosidase-producing non-Saccharomyces yeast strains were preliminarily screened by esculin medium, and the activities of β-glucosidase (β-Glu), α-L-arabinofuranosidase (α-L-Ara), α-L-rhamnosidase (α-L-Rha) and β-D-xylosidase (β-Xyl) originated from these screened strains were determined quantitatively by p-NP method. Comparing the glucosidase producing ability of non-Saccharomyces yeasts, the adaptability of the selected strains to wine fermentation conditions was analyzed. The results showed that non-Saccharomyces yeast autochthonous strains HX-1, HX-2, HX-3, and HX-4 had high glycosidase activity. According to the morphological characteristics of these strains grown on WL medium and the sequence analysis of 26S rDNA D1/D2 domain, HX-1 and HX-3 were identified as Hanseniaspora uvarum, while HX-2 and HX-4 were Metschnikowia pulcherrima. The individual glycosidase produced by these strains exhibited high activity at pH 5.0-6.0 and 40-50 ℃. In addition, high concentration of alcohol, SO2, and glucose can inhibit the glucosidase activity. The activity of β-Glu produced by HX-2 and HX-4 strains, α-L-Rha by HX-1 strain, α-L-Ara by HX-4 strain, and β-Xyl by HX-1 and HX-3 strains was high under the conditions of wine fermentation. Overall, our study provides technical support for the directional selection of the yeast strain with good aroma enhancement performance according to the type of glucosides in wine grapes.
[1] PADILLA B, GIL J V, MANZANARES P.Past and future of non-Saccharomyces yeasts:From spoilage microorganisms to biotechnological tools for improving wine aroma complexity[J].Frontiers in Microbiology, 2016, 7:411.
[2] 张慧敏, 潘秋红.葡萄酒中挥发性萜烯物质的产生机制及影响因素研究进展[J].食品科学, 2021, 42(13):249-258.
ZHANG H M, PAN Q H.Advances in understanding the formation mechanism of terpenoids during winemaking and factors influencing it[J].Food Science, 2021, 42(13):249-258.
[3] 杨旖旎. 武陵地区五种天然产物游离和键合风味成分的研究[D].恩施:湖北民族大学, 2019.
YANG Y N.Study on free and bound aroma components of five natural products in Wuling area[D].Enshi:Hubei Minzu University, 2019.
[4] 李蔚. 外源茉莉酸甲酯调控‘贵人香’葡萄品质机制研究[D].兰州:甘肃农业大学, 2020.
LI W.Mechanism of exogenous methyl jasmonate in regulating grape (Vitis vinifera L.cv.Italian Riesling) quality[D].Lanzhou:Gansu Agricultural University, 2020.
[5] HAMPEL D, ROBINSON A L, JOHNSON A J, et al.Direct hydrolysis and analysis of glycosidically bound aroma compounds in grapes and wines:Comparison of hydrolysis conditions and sample preparation methods[J].Australian Journal of Grape & Wine Research, 2014, 20(3):361-377.
[6] 刘玥姗. 葡萄果实糖苷酶活性、香气糖苷总量及其与成熟指标的关联分析[D].杨凌:西北农林科技大学, 2015.
LIU Y S.Correlation analysis of glucosidase activity, concentration of glycosidic aroma precursors and maturity indices of grape berries[D].Yangling:Northwest A & F University, 2015.
[7] 马得草. 优选非酿酒酵母糖苷酶的葡萄酒酿造环境适应性及其混菌发酵研究[D].杨凌:西北农林科技大学, 2017.
MA D C.The wine making environment adaptability of glycosidase from selected non-Saccharomyses and its mix-fermentation[D].Yangling:Northwest A & F University, 2017.
[8] 战吉宬, 曹梦竹, 游义琳, 等.非酿酒酵母在葡萄酒酿造中的应用[J].中国农业科学, 2020, 53(19):4057-4069.
ZHAN J C, CAO M Z, YOU Y L, et al.Research advance on the application of non-Saccharomyces in winemaking[J].Scientia Agricultura Sinica, 2020, 53(19):4057-4069.
[9] 彭传涛. 基于野生非酿酒酵母及其糖苷酶调控葡萄酒香气质量的研究[J].杨凌:西北农林科技大学, 2014.
PENG C T.Study on the enhancement of wine aroma using wild non-Sacchaeomyces and their glycosidase[J].Yangling:Northwest A & F University, 2014.
[10] 徐建坤. 产β-葡萄糖苷酶酵母菌的分离鉴定及特性研究[D].石河子:石河子大学, 2019.
XU J K.Isolation, identification and characterization of β-glucosidase producing yeast[D].Shihezi:Shihezi University, 2019.
[11] SPAGNA G, BARBAGALLO R.N, PALMERI R, et al.Properties of endogenous β-glucosidase of a Saccharomyces cerevisiae strain isolated from Sicilian musts and wines[J].Enzyme and Microbial Technology, 2002, 31(7): 1030-1035.
[12] SABEL A, MARTENS S, PETRI, A, et al.Wickerhamomyces anomalus AS1:A new strain with potential to improve wine aroma[J]. Annals of Microbiology, 2014, 64(2):483-491.
[13] 耿海波. 优良耐酸酿酒酵母的筛选及发酵特性研究[J].中国酿造, 2021, 40(10):152-156.
GENG H B.Screening and fermentation characteristics of superior acid-tolerant Saccharomyces cerevisiae[J].China Brewing, 2021, 40(10):152-156.
[14] 尹荐, 陶永胜, 孙玮璇, 等.优选非酿酒酵母胞外酶增香酿造干白葡萄酒效果[J].农业工程学报, 2020, 36(4):278-286.
YIN J, TAO Y S, SUN W X, et al.Effect of aroma enhancement for dry white wine by selected non-Saccharomyces extracellular enzymes[J].Transactions of the Chinese Society of Agricultural Engineering, 2020, 36(4):278-286.
[15] 马得草, 游灵, 李爱华, 等.高产β-葡萄糖苷酶野生酵母的快速筛选及其糖苷酶酿造适应性研究[J].西北农林科技大学学报(自然科学版), 2018, 46(1):129-135.
MA D C, YOU L, LI A H, et al.Rapid screen of wild yeast with high β-glucosidase activity and evaluation of its glycosidase in winemaking[J].Journal of Northwest A ＆ F University (Natural Science Edition), 2018, 46(1):129-135.
[16] 王佳, 胡兰兰, 张军翔, 等.高产β-葡萄糖苷酶野生酵母的筛选及产酶能力差异性分析[J].中国酿造, 2018, 37(2):50-53.
WANG J, HU L L, ZHANG J X, et al.Screening of wild yeast with high yield β-glycosidase and difference analysis of the enzyme production ability[J].China Brewing, 2018, 37(2):50-53.
[17] LÓPEZ M C, MATEO J J, MAICAS S.Screening of β-glucosidase and β-xylosidase activities in four non-Saccharomyces yeast isolates[J].Journal of Food Science, 2015, 80(8):1696-1704.
[18] 祝霞, 赵丹丹, 李俊娥, 等.O.oeni糖苷酶活性对干白葡萄酒萜烯类香气的影响[J].农业机械学报, 2021, 52(8):363-373.
ZHU X, ZHAO D D, LI J E, et al.Effect of glycosidase activity of Oenococcus oeni on terpene aroma compounds of dry white wine[J].Transactions of the Chinese Society for Agricultural Machinery, 2021, 52(8):363-373.
[19] MATEO J J, PERIS L, IBAÑEZ C, et al.Characterization of glycolytic activities from non-Saccharomyces yeasts isolated from Bobal musts[J].Journal of Industrial Microbiology & Biotechnology, 2011, 38(2):347-354.
[20] TESTA B, LOMBARDI S J, IORIZZO M, et al.Use of strain Hanseniaspora guilliermondii BF1 for winemaking process of white grapes Vitis vinifera cv Fiano[J].European Food Research & Technology, 2020, 246(3):549-561.
[21] KUO H P, WANG R B, HUANG C Y, et al.Characterization of an extracellular beta-glucosidase from Dekkera bruxellensis for resveratrol production[J].Journal of Food and Drug Analysis, 2018, 26(1):163-171.
[22] 朱晓琳. 糖苷酶处理葡萄酒酿造过程中香气物质的动态变化研究[D].杨凌:西北农林科技大学, 2017.
ZHU X L.Evolution of aroma compounds during winemaking process through the treatment with glycosidases[D].Yangling:Northwest A & F University, 2017.
[23] HU K, ZHU X L, MU H, et al.A novel extracellular glycosidase activity from Rhodotorula mucilaginosa:Its application potential in wine aroma enhancement[J].Letters in Applied Microbiology.2016, 62(2):169-176.
[24] BAFFI M A, TOBAL T, LAGO J H G, et al.Wine aroma improvement using a β-glucosidase preparation from Aureobasidium pullulans[J].Applied Biochemistry and Biotechnology, 2013, 169(2):493-501.
[25] PORTER T J, DIVOL B, SETATI M E.Lachancea yeast species:Origin, biochemical characteristics and oenological significance[J].Food Research International, 2019, 119:378-389.
[26] 刘浩. 葡萄模拟发酵体系中香气糖苷水解及发酵香气成分生成的动力学特征[D].杨凌:西北农林科技大学, 2016.
LIU H.Kinetic characteristics of hydrolysis of grape aroma glycosides and production of fermentative aromas during stimulated fermentation process[D].Yangling:Northwest A & F University, 2016.
[27] 问亚琴, 崔婧, 潘秋红.葡萄果实糖苷键合态萜烯物质的研究进展[J].园艺学报, 2012, 39(9):1679-1686.
WEN Y Q, CUI J, PAN Q H.Research progress of glycosidically-bound terpenes in grapes[J].Acta Horticulturae Sinica, 2012, 39(9):1679-1686.
[28] SONG H C, WANG X J, LI A H.et al. Profiling terpene glycosides from ecolly, cabernet gernischet, and Muscat hamburg grapes by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry[J].Journal of Food Science, 2020, 85(7):2032-2040.
[29] 刘吉彬. 葡萄萜烯类香气糖苷的质谱分析研究[D].杨凌:西北农林科技大学, 2017.
LIU J B.Mass spectrometric analysis of aromatic terpene glycosides in grape[D].Yangling:Northwest A & F University, 2017.
