乳酸菌发酵花生衣提取物过程中的代谢物特征分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-1300

摘要/Abstract

摘要：

目的探究保加利亚乳杆菌PS-1与嗜热链球菌PS-3发酵改善花生衣品质的代谢机制，解析其降低涩味、提升抗氧化活性的内在原因。方法以花生衣提取物（peanut skin extract, PSE）为原料，分别接种PS-1和PS-3菌株，采用非靶向代谢组学技术，结合多变量统计分析，对两种乳酸菌在不同时间点（0、12、24、36、48 h）发酵产物进行检测，筛选显著差异代谢物，并分析其通路富集与变化趋势。结果乳酸菌发酵能显著改变花生衣提取物的代谢谱，保加利亚乳杆菌和嗜热链球菌分别筛选到49种和88种显著差异代谢物；KEGG分析表明，代谢物显著富集于酪氨酸代谢、黄酮类降解与生物合成及嘌呤代谢等通路；发酵实现了酚类物质的生物转化，保加利亚乳杆菌发酵过程中，二氢杨梅素、原花青素B2及二氢山柰酚等含量显著降低，而辛弗林、1-脱氧野尻霉素、富马酸及丙酮酸等含量显著增加；嗜热链球菌发酵中，白藜芦醇、原花青素A2、黄芩素及槲皮素等物质含量下降，棉花皮苷、木犀草素苷及异荭草素等物质含量显著上升；两种菌对酚类物质的生物转化作用均发生在发酵12-24 h段。结论乳酸菌发酵实现花生衣酚类物质的生物转化，促进了抗氧化、减脂及降糖等相关功能代谢物的积累，同时有效降低了潜在涩味物质含量。

关键词: 乳酸菌, 花生衣提取物, 发酵过程, 非靶向代谢组学, 酚类物质

Abstract:

Objective To investigate the metabolic mechanisms of the quality improvement of peanut skin by fermentation with Lactobacillus delbrueckii subsp. bulgaricus PS-1 and Streptococcus thermophilus PS-3, and to elucidate the underlying reasons for the reduction of astringency and the enhancement of antioxidant activity. Method Peanut skin extract (PSE) was fermented using strains PS-1 and PS-3, respectively. Fermentation products were collected at different time points (0, 12, 24, 36, and 48 h) and analyzed using untargeted metabolomics techniques combined with multivariate statistical analysis. Significantly differential metabolites were screened, followed by analyzing their pathway enrichment and temporal variation trends. Result Fermentation by lactic acid bacteria significantly altered the metabolic profile of PSE. Compared to the unfermented control, 49 and 88 significantly differential metabolites were identified in PSE fermented by Lactobacillus bulgaricus PS-1 and Streptococcus thermophilus PS-3, respectively. KEGG pathway analysis revealed significant enrichment in tyrosine metabolism, flavonoid biosynthesis and degradation, and purine metabolism. Fermentation facilitated the biotransformation of phenolic compounds. During L. bulgaricus PS-1 fermentation, the contents of dihydromyricetin, procyanidin B2, and dihydrokaempferol decreased significantly, while the levels of synephrine, 1-deoxynojirimycin, fumaric acid, and pyruvic acid increased markedly. In S. thermophilus PS-3 fermentation, the contents of resveratrol, procyanidin A2, baicalein, and quercetin declined, whereas the levels of gossypin, luteolin glycoside, and isoorientin rose significantly. The biotransformation of phenolic substances by both strains primarily occurred during the 12-24 h fermentation period. Conclusion Fermentation by lactic acid bacteria facilitates the biotransformation of phenolic compounds in peanut skins, promoting the accumulation of functional metabolites with antioxidant, lipid-lowering, and hypoglycemic activities, and resulting in reduced levels of potential astringent substances.

Key words: lactic acid bacteria, peanut skin extract, fermentation process, untargeted metabolomics, phenolic compounds

张初署, 曹世宁, 王发明, 周海翔, 胡晓君, 唐月异, 周晓燕, 王冕, 陈静, 张建成. 乳酸菌发酵花生衣提取物过程中的代谢物特征分析[J]. 生物技术通报, 2026, 42(5): 340-352.

ZHANG Chu-shu, CAO Shi-ning, WANG Fa-ming, ZHOU Hai-xiang, HU Xiao-jun, TANG Yue-yi, ZHOU Xiao-yan, WANG Mian, CHEN Jing, ZHANG Jian-cheng. Analysis of Metabolite Characteristics during Lactic Acid Bacteria Fermentation of Peanut Skin Extract[J]. Biotechnology Bulletin, 2026, 42(5): 340-352.

图/表 8

图1 不同发酵阶段发酵液的PLS-DA图A：保加利亚乳杆菌PS-1；B：嗜热链球菌PS-3；B-0 h、B-12 h、B-24 h、B-36 h、B-48 h表示保加利亚乳杆菌PS-1发酵0、12、24、36、48 h；S-0 h、S-12 h、S-24 h、S-36 h、S-48 h表示嗜热链球菌PS-3发酵0、12、24、36、48 h

Fig. 1 PLS-DA plot of fermentation broths at different fermentation stagesA: Lactobacillus bulgaricus PS-1. B: Streptococcus thermophilus PS-3;B-0, B-12 h, B-24 h, B-36 h, and B-48 h refer to the fermentation time points (0, 12, 24, 36, and 48 h) of L. bulgaricus PS-1; S-0 h, S-12 h, S-24 h, S-36 h, and S-48 h indicatethe fermentation time points (0, 12, 24, 36, and 48 h) of S. thermophilus PS-3

表1 保加利亚乳杆菌PS-1发酵前后差异代谢物种类与变化情况

Table 1 Categories and changes of differential metabolites before and after fermentation with L. bulgaricus PS-1

代谢物类别 Metabolite class （Super class）	显著差异代谢物 Number of significantly differential metabolites	占比 Proportion （%）	代谢物类别 Metabolite class	上调 Up-regulated	下调 Down-regulated
有机杂环化合物	12	24.49	吲哚及其衍生物	3	0
			嘌呤核苷	0	2
			哌啶类	2	0
			苯并恶嗪	1	0
			羟基酸及其衍生物	0	1
			喹啉及其衍生物	1	0
			内酯	0	1
苯丙烷和聚酮	9	18.37	黄酮类化合物	1	4
			香豆素及其衍生物	2	0
			肉桂酸及其衍生物	1	0
			异黄酮类	0	1
有机酸及其衍生物	8	16.33	羧酸及其衍生物	5	1
			鞘磷脂	1	0
			有机膦酸及其衍生物	0	1
脂质和类脂分子	7	14.29	脂肪酰基	0	4
			酮酸及其衍生物	1	1
			类固醇和类固醇衍生物	0	1
苯环型化合物	7	14.29	苯及其取代衍生物	2	3
			酚类	1	1
有机含氧化合物	5	10.20	有机氧化合物	2	3
核苷、核苷酸和类似物	1	2.04	嘌呤核苷酸	1	0

表2 嗜热链球菌PS-3发酵前后差异代谢物种类与变化情况

Table 2 Categories and changes of differential metabolites before and after fermentation with S. thermophilus PS-3

代谢物类别 Metabolite class （Super class）	显著差异代谢物 Number of significantly differential metabolites	占比 Proportion （%）	代谢物类别 Metabolite class	上调 Up-regulated	下调 Down-regulated
苯丙烷和聚酮	23	26.14	黄酮类化合物	8	11
			肉桂酸及其衍生物	2	0
			香豆素及其衍生物	0	1
			芪类	0	1
有机含氧化合物	14	15.91	有机氧化合物	5	9
有机杂环化合物	14	15.91	苯骈吡喃	2	1
			二氮杂苯	1	1
			吡啶及其衍生物	1	1
			吲哚及其衍生物	1	0
			噻吩	0	1
			内酯	0	1
			咪唑并嘧啶类	0	1
			恶嗪类	0	1
			吡咯吡嗪类	0	1
			苯并噻嗪类	0	1
有机酸及其衍生物	11	12.50	羧酸及其衍生物	1	7
			有机硫酸及其衍生物	1	0
			有机硫代磷酸及其衍生物	1	0
苯环型化合物	10	11.36	酮酸及其衍生物	0	1
			酚类	1	1
			萘类	1	0
脂质和类脂分子	9	10.23	脂肪酰基	1	5
			甘油磷酯	0	2
			类固醇和类固醇衍生物	0	1
核苷、核苷酸和类似物	4	4.55	嘌呤核苷酸	1	0
			嘌呤核苷	0	1
			吡啶核苷酸	0	1
			（3'->5'）-二核苷酸和类似物	1	0
有机氮化合物	2	2.27	有机氮化合物	1	1
生物碱及其衍生物	1	1.14	哈马拉生物碱	0	1

图2 保加利亚乳杆菌PS-1发酵过程中主要通路的化合物变化趋势A：芳香族化合物降解通路；B：类黄酮化合物的降解与合成通路；C：酪氨酸代谢通路；D：嘌呤代谢通路。Lg（S）表示峰面积的对数值（以10为底）

Fig. 2 Change trends of compounds in the key pathways during L. bulgaricus PS-1 fermentationA: Aromatic compound degradation pathway. B: Flavonoid biosynthesis and degradation pathway. C: Tyrosine metabolism pathway. D: Purine metabolism pathway. Lg(S) indicates the logarithm of the peak area (with base 10)

图3 嗜热链球菌PS-3发酵过程中主要代谢通路中化合物变化趋势A：核苷酸糖生物合成通路；B：类黄酮化合物降解/合成通路；C：酪氨酸代谢通路；lg（S）表示峰面积的对数值（以10为底）

Fig. 3 Change trends of compounds in the key pathways during S. thermophilus PS-3 fermentationA: Nucleotide sugar biosynthesis pathway. B: Flavonoid biosynthesis and degradation pathway. C: Tyrosine metabolism pathway. lg(S) indicates the logarithm of the peak area (with base 10)

图4 乳酸菌发酵过程中代谢物含量变化热图A：保加利亚乳杆菌PS-1；B：嗜热链球菌PS-3

Fig. 4 Heat map of metabolite changes during lactic acid bacteria fermentationA: L. bulgaricus PS-1; B: S. thermophilus PS-3

图5 保加利亚乳杆PS-1发酵花生衣提取物过程中代谢网络分析COMT: Catechol-O-methyltransferase. ADH: Alcohol dehydrogenase. ALDH: Aldehyde dehydrogenase. PcaF: Protocatechuate 3,4-dioxygenase (beta subunit). PK: Pyruvate kinase. PDHc: Pyruvate dehydrogenase complex. ACC: Acetyl-CoA carboxylase. 4CL: 4-Coumarate: CoA ligase. PEPC: Phosphoenolpyruvate carboxylase. PC: Pyruvate carboxylase. MDH: Malate dehydrogenase. FH: Fumarate hydratase. FRD: Fumarate reductase. CHS: Chalcone synthase. CHI: Chalcone isomerase. F3H: Flavanone 3-hydroxylase. DFR: Dihydroflavonol 4-reductase. FLS: Flavonol synthase. ANS: Anthocyanidin synthase. UGT: UDP-glucose: Flavonoid glycosyltransferase. ANR: Anthocyanidin reductase. OC: Oxidase/Cyclase. TDC: Tyrosine decarboxylase. DBH: Dopamine beta-hydroxylase. PNMT: Phenylethanolamine N-methyltransferase. TAL: Tyrosine ammonia-lyase. TAT: Tyrosine aminotransferase. PPDC: 4-Hydroxyphenylpyruvate dioxygenase. MHPG: 3-methoxy-4-hydroxyphenylglycol. DHPG: 3,4-Dihydroxyphenylglycol. DOPGAL: 3,4-Dihydroxyphenylglycolaldehyde. DHMA: 3,4-Dihydroxymandelic acid. HPP: 4-Hydroxyphenylpyruvate. HPAL: 4-Hydroxyphenylacetaldehyde

Fig. 5 Metabolic network analysis during the fermentation of peanut skin extract by L. bulgaricus PS-1

图6 嗜热链球菌PS-3发酵花生衣提取物过程中代谢网络分析AT: AcylTransferase. pAMT: Putative aminotransferase. PAL: Phenylalanine ammonia-lyase. CHS: Chalcone synthase. ALDH: Aldehyde dehydrogenase. COMT: Catechol-O-methyltransferase. CHI: Chalcone isomerase. F3H: Flavanone 3-hydroxylase. F3'H: Flavonoid 3'-hydroxylase. FLS: Flavonol synthase. F3'5'H: Flavonoid 3',5'-hydroxylase. ANS: Anthocyanidin synthase. ANR: Anthocyanidin reductase. OC: Oxidase/Cyclase. DFR: Dihydroflavonol 4-reductase. F8H: Flavonoid 8-hydroxylase. UGT: UDP-glycosyltransferase. FNR: Flavonoid NADPH reductase. FNS: Flavone synthase. C-GT: C-glycosylTransferase. OMT: O-Methyltransferase. SSH: Scutellaria-specific hydroxylase. F6H: Flavone 6-hydroxylase. DHMA: 3,4-Dihydroxymandelic acid. DOPGAL: 3,4-Dihydroxyphenylglycolaldehyde

Fig. 6 Metabolic network analysis during the fermentation of peanut skin extract by S. thermophilus PS-3

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