Functional peptidomics analysis of Saccharomyces pastorianus protein hydrolysates based on different enzyme treatments

doi:10.3724/SP.J.1123.2023.08029

Abstract

Abstract:

The aim of this study is to explore differences in the peptidomics of Saccharomyces pastorianus protein hydrolysates treated with different enzymes. Briefly, differences in the peptide fingerprints and active peptides of neutral protease/papain-hydrolyzed S. pastorianus were analyzed using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) combined with PEAKS Online 1.7 analysis software, Peptide Ranker, and the BIOPEP database. Compared to traditional databases, the PEAKS Online uses de novo sequencing for analysis to obtain oligopeptides smaller than pentapeptides. It provides more comprehensive data of the peptide sample. In this study, enzymatic hydrolysates of S. pastorianus protein were prepared under the optimum conditions of neutral protease and papain respectively. In total, 7221 and 7062 polypeptides were identified in the hydrolysates of neutral protease and papain, respectively; among these polypeptides, 980 were common to the two enzymes. The 6241 and 6082 unique peptides found in the hydrolysates of neutral protease and papain, respectively, indicated that the peptide fingerprints of the two hydrolysates are quite different. Peptide Ranker predicted that 3013 (41.73%) and 3095 (43.83%) peptides were potentially bioactive in the hydrolysates of neutral protease and papain, respectively. According to the BIOPEP database, neutral protease and papain contained 295 and 357 active peptides, respectively; these peptides were mainly composed of angiotensin converting enzyme (ACE) inhibitors and dipeptidyl peptidase IV inhibitors and antioxidant peptides. The number of active peptides in the hydrolysate of papain was higher than that in the hydrolysate of neutral protease, but the total ion intensity of active peptides in the former was lower than that in the latter. This study revealed the influence of protease type on the composition of enzymatic hydrolysates from S. pastorianus protein. The above results provide a reference for the development of functional products of S. pastorianus protein peptides and the high-value utilization of yeast resources.

Key words: ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS), peptide fingerprint, active peptide, Saccharomyces pastorianus

CLC Number:

O658

YAN Yutong, GAO Chunyu, ZHANG Xiaomei, AN Zizhe, MA Yunzhen, HAN Linlin, ZHANG Hongwei, ZHAO Xue. Functional peptidomics analysis of Saccharomyces pastorianus protein hydrolysates based on different enzyme treatments[J]. Chinese Journal of Chromatography, 2023, 41(11): 995-1001.

Figures/Tables 5

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Peptide composition	Number of common peptides	Specific peptides
		Neutral protease		Papain
		Number	Relative abundance/%	Number	Relative abundance/%
Amino acid	9	0	1.36	3	5.76
Dipeptide	90	25	30.78	38	22.62
Tripeptide	309	231	18.67	474	28.41
Tetrapeptide	384	1120	21.98	1883	26.90
Pentapeptide	171	1987	13.32	2109	11.62
Hexapeptide	17	1204	6.46	873	2.26
Heptapeptide	6	681	3.39	337	0.84
Octapeptide	2	447	1.82	148	0.72
Nonapeptide	1	206	0.67	79	0.17
Decapeptide	0	133	0.38	43	0.22
More than decapeptide	0	207	1.18	98	0.48
Total	980	6241		6082

Peptide composition	Number of common peptides	Specific peptides
		Neutral protease		Papain
		Number	Relative abundance/%	Number	Relative abundance/%
Amino acid	9	0	1.36	3	5.76
Dipeptide	90	25	30.78	38	22.62
Tripeptide	309	231	18.67	474	28.41
Tetrapeptide	384	1120	21.98	1883	26.90
Pentapeptide	171	1987	13.32	2109	11.62
Hexapeptide	17	1204	6.46	873	2.26
Heptapeptide	6	681	3.39	337	0.84
Octapeptide	2	447	1.82	148	0.72
Nonapeptide	1	206	0.67	79	0.17
Decapeptide	0	133	0.38	43	0.22
More than decapeptide	0	207	1.18	98	0.48
Total	980	6241		6082

Type	No.	Sequence	Score		z	Relative abundance/%
Neutral protease	1	FFPF	0.9981	557.2792	1	0.0027
	2	FGFWF	0.9981	352.1639	2	0.1091
	3	WWGF	0.9978	595.2611	1	0.0002
	4	WMF	0.9976	483.2065	1	0.0005
	5	WMW	0.9972	522.2168	1	0.0010
	6	WFP	0.9971	449.2188	1	0.0013
	7	WGF	0.9969	409.1870	1	0.0015
	8	PFF	0.9968	410.2078	1	0.0014
	9	FLFF	0.9965	573.3081	1	0.0035
	10	FPMF	0.9965	541.2482	1	0.0002
Papain	1	WFW	0.9988	538.2451	1	0.0050
	2	FMF	0.9977	444.1951	1	0.0022
	3	FPW	0.9971	449.2184	1	0.0056
	4	GFW	0.9970	409.1874	1	0.0040
	5	GFF	0.9970	370.1764	1	0.0213
	6	WPW	0.9968	488.2296	1	0.0015
	7	WLFW	0.9964	651.3292	1	0.0039
	8	FFPGW	0.9963	653.3026	1	0.0001
	9	WDFF	0.9960	614.2609	1	0.0064
	10	FFPM	0.9960	541.2505	1	0.0037

Type	No.	Sequence	Score		z	Relative abundance/%
Neutral protease	1	FFPF	0.9981	557.2792	1	0.0027
	2	FGFWF	0.9981	352.1639	2	0.1091
	3	WWGF	0.9978	595.2611	1	0.0002
	4	WMF	0.9976	483.2065	1	0.0005
	5	WMW	0.9972	522.2168	1	0.0010
	6	WFP	0.9971	449.2188	1	0.0013
	7	WGF	0.9969	409.1870	1	0.0015
	8	PFF	0.9968	410.2078	1	0.0014
	9	FLFF	0.9965	573.3081	1	0.0035
	10	FPMF	0.9965	541.2482	1	0.0002
Papain	1	WFW	0.9988	538.2451	1	0.0050
	2	FMF	0.9977	444.1951	1	0.0022
	3	FPW	0.9971	449.2184	1	0.0056
	4	GFW	0.9970	409.1874	1	0.0040
	5	GFF	0.9970	370.1764	1	0.0213
	6	WPW	0.9968	488.2296	1	0.0015
	7	WLFW	0.9964	651.3292	1	0.0039
	8	FFPGW	0.9963	653.3026	1	0.0001
	9	WDFF	0.9960	614.2609	1	0.0064
	10	FFPM	0.9960	541.2505	1	0.0037