刺葡萄酒酒精发酵过程中氨基酸组成与果香酯类物质 生成的关联分析

doi:10.3864/j.issn.0578-1752.2020.11.013

摘要/Abstract

摘要：

【目的】研究刺葡萄发酵过程中氨基酸的化学组成与果香酯类物质生成的关联,为刺葡萄酒增香酿造提供理论指导。【方法】试验以刺葡萄品种‘湘珍珠’为研究对象,以‘赤霞珠’葡萄为对照,添加酿酒酵母启动酒精发酵,每隔24 h取样,分别采用HPLC和GC-MS方法测定氨基酸和酯类物质的组成及含量,并通过Pearson相关性分析建立二者之间的数学关联。【结果】刺葡萄中除脯氨酸之外的氨基酸总体含量为‘赤霞珠’葡萄的3倍,天冬氨酸、缬氨酸、谷氨酸、γ-氨基丁酸、精氨酸含量分别达到‘赤霞珠’葡萄中对应氨基酸含量的2.4倍、2.2倍、4.2倍、7.7倍和3倍,但丙氨酸、苯丙氨酸、异亮氨酸含量仅为‘赤霞珠’葡萄中对应氨基酸含量的10%、35%和20%。刺葡萄发酵产生的乙酸酯、短链脂肪酸乙酯、中链脂肪酸乙酯含量仅为‘赤霞珠’葡萄发酵所得上述酯类物质的55%、11%和69%。Pearson相关性分析显示,刺葡萄发酵过程中丙氨酸、苯丙氨酸、异亮氨酸与乙酸酯、脂肪酸乙酯的相关性系数在0.552—0.837,较‘赤霞珠’葡萄发酵体系中相应的相关性系数显著偏低,这可能与刺葡萄酒果香酯类物质含量偏低有关。【结论】刺葡萄中含量偏低的丙氨酸、苯丙氨酸、异亮氨酸可能限制了刺葡萄酒发酵体系中果香酯类物质的积累。

关键词: 葡萄酒, 刺葡萄, 氨基酸, 果香酯类, 相关性分析

Abstract:

【Objective】The aim of this study was to reveal the correlation between amino acid composition and fruity ester production during fermentation of spine grapes, so as to provide the theoretical guidance for aroma enhancement fermentation of spine grape wine. 【Method】A spine grape variety, Xiangzhenzhu, was used to conduct alcoholic fermentation with Saccharomyces cerevisiae, and Cabernet Sauvignon grape was served as control. During fermentation, sampling was carried out every 24 h to analyze the concentrations of amino acids and esters by using HPLC and GC-MS, respectively. Furthermore, the mathematical relationship between these two parameters was analyzed by Pearson correlation analysis. 【Result】Total content of amino acids (except proline) in spine grape was 3 folds higher than that in Cabernet Sauvignon grape, with 2.4, 2.2, 4.2, 7.7 and 3 folds for aspartic acid, valine, glutamic acid, γ-aminobutyric acid and arginine, respectively. However, the content of alanine, phenylalanine and isoleucine in spine grape only accounted for 10%, 35% and 20%, respectively. Compared with Cabernet Sauvignon, fermentation with spine grape only produced 55%, 11% and 69% in the concentrations of acetates, short-chain fatty acid ethyl esters, and medium-chain fatty acid ethyl esters, respectively. Pearson correlation analysis further showed that the correlation coefficient between alanine, phenylalanine, isoleucine and acetate, fatty acid ethyl ester was 0.552-0.837 during fermentation with spine grape, which was significantly lower than that of Cabernet Sauvignon. This might be related to low content of fruity esters in spine grape wine. 【Conclusion】Low content of alanine, phenylalanine and isoleucine could negatively affect ester productions during alcoholic fermentation.

Key words: wine, spine grape, amino acid, fruity ester, correlation analysis

朱子健,陈思雨,粟俊,陶永胜. 刺葡萄酒酒精发酵过程中氨基酸组成与果香酯类物质生成的关联分析[J]. 中国农业科学, 2020, 53(11): 2272-2284.

ZHU ZiJian,CHEN SiYu,SU Jun,TAO YongSheng. Correlation Analysis Between Amino Acids and Fruity Esters During Spine Grape Fermentation[J]. Scientia Agricultura Sinica, 2020, 53(11): 2272-2284.

图/表 6

表1

图1

表2

刺葡萄酒精发酵过程中酯类物质的含量"

化合物 Compound		葡萄醪 Must	1 d	2 d	3 d	5 d	7 d	10 d	12 d	嗅觉阈值 Threshold (μg?L^-1)	气味描述 Odor description
乙酸酯 Acetate		2352.3±0.1	1861.9±17.6	2914.9±105.4	3608.9±7.7	8826.9±567.5	11450.5±1056.8	13251.9±499.8	16233.0±129.2
1	乙酸乙酯 Ethyl acetate	2352.3±0.1	1861.9±17.6	2913.8±105.5	3608.1±7.7	8644.5±526.1	11383.7±1051.5	13204.6±499.2	16168.6±117.5	7500^[4]	菠萝,果香,甜香 Pineapple, fruity, sweet
2	乙酸异丁酯 Isobutyl acetate	ND	ND	ND	ND	18.7±0.3	7.4±0.5	ND	0.4±0.1	1600^[4]	果香,梨,香蕉 Fruity, pear, banana
3	乙酸异戊酯Isoamyl acetate	ND	ND	ND	ND	161.4±41.5	57.6±5.5	46.0±0.5	63.1±11.7	30^[17]	果香,香蕉Fruity, banana
4	乙酸丁酯 Butyl acetate	ND	ND	1.0±0.1	0.8±0.1	1.6±0.2	1.3±0.2	1.1±0.1	1.1±0.2	1600^[4]	果香,梨,香蕉 Fruity, pear, banana
5	乙酸己酯Hexyl acetate	ND	ND	ND	ND	ND	ND	ND	ND	1500^[17]	梨,甜果Pear, sweet fruit
6	乙酸辛酯Octyl acetate	ND	ND	ND	ND	0.7±0.1	0.4±0.2	0.3±0.1	ND	NF
7	2-乙酸苯乙酯 2-phenylethyl acetate	ND	ND	ND	8.1±0.1	12.3±0.4	17.4±2.0	20.4±0.9	20.9±0.1	250^[18]	花香Floral
短链脂肪酸乙酯 SCFAEEs		ND	ND	ND	2.6±0.2	66.2±8.0	90.3±4.8	106.1±6.5	132.4±3.6
1	丁酸乙酯 Ethyl butyrate	ND	ND	ND	ND	59.2±7.8	85.0±4.5	101.8±6.4	127.5±3.3	20^[4]	草莓,苹果,香蕉 Strawberry, apple, banana
2	异戊酸乙酯 Ethyl isovalerate	ND	ND	ND	2.6±0.2	7.0±0.2	5.3±0.3	4.3±0.1	4.9±0.3	3^[19]	香蕉,甜果 Banana, sweet fruity
中链脂肪酸乙酯 MCFAEEs		ND	ND	35.2±1.0	231.7±23.8	1263.0±99.7	933.1±33.4	711.4±22.7	711.9±13.5
1	己酸乙酯 Ethyl hexanoate	ND	ND	ND	73.6±12.5	707.3±73.7	531.9±11.8	414.9±21.5	429.2±9.3	5^[20]	花香,果香,草莓 Floral, fruity, strawberry
2	庚酸乙酯Ethyl heptanoate	ND	ND	ND	ND	0.9±0.1	0.7±0.3	0.6±0.1	0.5±0.1	220^[21]	白兰地,果香Brandy, fruity
3	辛酸乙酯 Ethyl octanoate	ND	ND	35.0±1.0	132.9±10.0	485.3±22.1	373.2±18.6	295.6±1.2	282.0±4.2	5^[4]	苹果,梨,花香 Apple, pear, floral
4	癸酸乙酯 Ethyl caprate	ND	ND	0.2±0.1	0.4±0.1	0.8±0.1	0.5±0.1	0.3±0.1	0.2±0.1	200^[4]	蜡味,果香,玫瑰 Waxy, fruity, rose
5	月桂酸乙酯 Ethyl dodecanoate	ND	ND	ND	24.8±1.3	68.7±3.9	26.8±2.6	ND	ND	1500^[19]	甜香,花香,果香,奶油 Sweet, floral, fruity, cream
其他酯类other esters		ND	ND	6.3±0.3	8.0±0.2	65.1±2.2	33.5±3.7	11.7±0.3	6.9±0.1
1	己酸甲酯Methyl caproate	ND	ND	6.3±0.3	7.9±0.1	44.1±0.7	17.9±1.4	ND	ND	NF
2	己酸异戊酯Isoamyl hexanoate	ND	ND	ND	0.1±0.1	6.5±0.5	4.0±0.2	2.4±0.1	1.5±0.1	NF	苹果,菠萝Apple, pineapple
3	辛酸异戊酯Isoamyl octanoate	ND	ND	ND	ND	14.5±1.0	11.6±2.1	9.3±0.2	5.4±0.1	125^[22]	甜香Sweat

表2

表3

赤霞珠酒精发酵过程中酯类物质的含量"

化合物 Compound		发酵日程 Fermentation time								嗅觉阈值 Threshold (μg?L^-1)	气味描述 Odor description
化合物 Compound		葡萄醪Must	1 d	2 d	3 d	5 d	7 d	10 d	12 d	嗅觉阈值 Threshold (μg?L^-1)	气味描述 Odor description
乙酸酯Acetate		1846.1±2.5	1951.8±35.4	3447.1±240.9	9036.5±1418.0	26065.4±1197.7	26151.0±105.7	28865.8±594.1	29631.1±1381.0
1	乙酸乙酯 Ethyl acetate	1846.0±2.5	1951.8±35.4	3437.4±240.8	8608.2±1403.7	24509.4±827.1	25681.0±69.4	26713.2±508.6	27764.9±1252.0	7500^[4]	菠萝,果香,甜香 Pineapple, fruity, sweet
2	乙酸异丁酯 Isobutyl acetate	ND	ND	ND	59.5±12.5	100.4±11.4	62.0±2.6	37.1±9.0	0.8±0.1	1600^[4]	果香,梨,香蕉 Fruity, pear, banana
3	乙酸异戊酯 Isoamyl acetate	ND	ND	ND	345.2±0.1	1383.3±355.5	306.7±19.0	1994.3±74.9	1753.5±127.5	30^[17]	果香,香蕉 Fruity, banana
4	乙酸丁酯 Butyl acetate	0.1±0.1	ND	2.1±0.1	5.0±0.6	8.3±0.5	8.3±0.1	8.3±0.2	6.4±0.3	1600^[4]	果香,梨,香蕉 Fruity, pear, banana
5	乙酸己酯 Hexyl acetate	ND	ND	ND	ND	ND	0.4±0.1	0.7±0.1	0.6±0.1	1500^[17]	梨,甜果 Pear, sweet fruit
6	乙酸辛酯 Octyl acetate	ND	ND	ND	0.3±0.1	0.8±0.1	0.6±0.1	0.4±0.1	0.3±0.1	NF
7	2-乙酸苯乙酯 2-phenylethyl acetate	ND	ND	7.6±0.1	18.3±1.1	63.2±3.1	92.0±14.6	111.8±1.4	104.6±1.2	250^[18]	花香 Floral
短链脂肪酸乙酯 SCFAEEs		ND	ND	12.7±0.1	752.2±146.9	2293.4±167.3	1683.0±79.6	1279.5±30.0	1253.4±83.5
1	丁酸乙酯 Ethyl butyrate	ND	ND	ND	28.9±6.7	111.7±5.3	127.3±4.2	138±5.0	143.6±10.0	20^[4]	草莓,苹果,香蕉 Strawberry, apple, banana
2	异戊酸乙酯 Ethyl isovalerate	ND	ND	ND	440.2±92.8	1146.5±77.0	841.7±14.9	638.8±19.3	530.7±40.5	3^[19]	香蕉,甜果 Banana, sweet fruity
中链脂肪酸乙酯 MCFAEEs		ND	ND	12.1±0.5	715.3±139.4	2149.7±159.3	1492.9±95.3	1058.5±27.2	1038.0±69.7
1	己酸乙酯 Ethyl hexanoate	ND	ND	ND	440.2±92.8	1146.5±77.0	841.7±14.9	638.8±19.3	530.7±40.5	5^[20]	花香,果香,草莓 Floral, fruity, strawberry
2	庚酸乙酯 Ethyl heptanoate	ND	ND	0.2±0.1	0.8±0.1	3.2±0.2	2.4±0.3	1.9±0.1	2.1±0.2	220^[21]	白兰地,果香 Brandy, fruity
3	辛酸乙酯 Ethyl octanoate	ND	ND	11.9±0.5	238.8±33.6	846.9±74.2	587.5±68.1	417.6±7.9	505.0±29.0	5^[4]	苹果,梨,花香 Apple, pear, floral
4	癸酸乙酯 Ethyl caprate	ND	ND	ND	0.6±0.1	0.8±0.1	0.5±0.1	0.2±0.1	0.2±0.1	200^[4]	蜡味,果香,玫瑰 Waxy, fruity, rose
5	月桂酸乙酯 Ethyl dodecanoate	ND	ND	ND	34.9±12.8	152.3±7.8	60.8±11.9	ND	ND	1500^[19]	甜香,花香,果香,奶油 Sweet, floral, fruity, cream
其他酯类 other esters		ND	ND	14.4±0.5	31.6±4.4	31.5±3.5	13.8±5.4	6.9±0.2	9.0±0.2
1	己酸甲酯Methyl caproate	ND	ND	14.2±0.5	25.8±3.4	6.8±1.5	2.0±1.2	ND	ND	NF
2	己酸异戊酯 Isoamyl hexanoate	ND	ND	0.2±0.1	1.4±0.3	7.7±0.7	4.0±0.7	1.5±0.2	1.7±0.3	NF	苹果,菠萝 Apple, pineapple
3	辛酸异戊酯 Isoamyl octanoate	ND	ND	ND	4.4±0.7	17.0±1.3	7.8±3.5	5.4±0.2	7.3±0.1	125^[22]	甜香 Sweat

表3

表4

图2

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氨基酸 Amino acid	赤霞珠葡萄 Cabernet Sauvignon grape	刺葡萄 Spine grape
天冬氨酸 Asp	11.7±2.6b	28.1±4a
谷氨酸 Glu	14.9±0.2b	63.4±18a
精氨酸 Arg	273.2±1.5b	814.7±33.8a
脯氨酸 Pro	10868.6±108a	1316.4±79.4b
γ-氨基丁酸 GABA	59.9±0.5b	463±3.2a
丙氨酸 Ala	48.1±4.4a	5±0.1b
缬氨酸 Val	14.3±0b	31.6±0.6a
蛋氨酸 Met	ND	ND
异亮氨酸 Ile	37.6±2.5a	7.56±0.5b
亮氨酸 Leu	ND	ND
色氨酸 Trp	ND	ND
苯丙氨酸 Phe	15.4±0.9a	5.4±1b
酪氨酸 Tyr	21.1±0.4a	16.9±1.2ab
总氨基酸 Total amino acids	11350.4±100a	3727.8±59.8b
总氨基酸（除脯氨酸） Total amino acids (except Pro)	481.8±11b	2411.4±52.9a

品种 Variety	化合物 Compound	总氨基酸 Total amino acid	天冬氨酸 Asp	谷氨酸 Glu	精氨酸 Arg	γ-氨基丁酸 GABA	缬氨酸 Val	丙氨酸 Ala	异亮氨酸 Ile	苯丙氨酸 Phe
赤霞珠 Cabernet Sauvignon	乙酸酯 Acetates	-0.919	-0.865	-0.810	-0.910	-0.848	-0.750	-0.946	-0.764	-0.978
	短链脂肪酸乙酯 SCFAEEs	-0.875	-0.831	-0.754	-0.858	-0.791	-0.695	-0.961	-0.705	-0.945
	中链脂肪酸乙酯 MCFAEEs	-0.825	-0.736	-0.762	-0.833	-0.796	-0.689	-0.733	-0.716	-0.876
	其他酯类 Other esters	-0.505	-0.419	-0.612	-0.541	-0.593	-0.591	-0.139	-0.607	-0.378
刺葡萄 Spine grape	乙酸酯 Acetates	-0.873	-0.927	-0.889	-0.888	-0.976	-0.821	-0.837	-0.738	-0.694
	短链脂肪酸乙酯 SCFAEEs	-0.864	-0.929	-0.913	-0.886	-0.970	-0.759	-0.805	-0.791	-0.661
	中链脂肪酸乙酯 MCFAEEs	-0.750	-0.721	-0.892	-0.869	-0.745	-0.669	-0.794	-0.552	-0.569
	其他酯类 Other esters	-0.399	-0.331	-0.571	-0.558	-0.298	-0.500	-0.370	-0.178	-0.216