Optimization of Hydrolysis and Identifcation of Bioactive Peptides in Brewery Yeast Residuals

Rosa, Fernanda A. Della; Tonin, Angelica P.; Rocha, Beatriz S.; Santos, Marco A. R.; Silveira, Fabiane M.; Cardoso-Filho, Lucio; Ribeiro, Valquíria M. S.; Meurer, Eduardo C.

doi:10.21577/0103-5053.20230146

Fernanda A. Della Rosa

investigation

writing de original draft

Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Laboratório Fenn de Espectrometria de Massas, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

https://orcid.org/0000-0002-0853-2488

Angelica P. Tonin

data analysis

Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Laboratório Fenn de Espectrometria de Massas, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Beatriz S. Rocha

analysis

Laboratório Fenn de Espectrometria de Massas, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Marco A. R. Santos

experimental design

data analysis

Laboratório Fenn de Espectrometria de Massas, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Fabiane M. Silveira

data analysis

Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Lucio Cardoso-Filho

conceptualization

experimental design

Departamento de Engenharia Química, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Valquíria M. S. Ribeiro

validation

manuscript review

Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Laboratório Fenn de Espectrometria de Massas, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Eduardo C. Meurer ^** e-mail: eduardo.meurer@ufpr.br

funding acquisition

validation

manuscript writing, review and editing

experimental design

supervision

Departamento de Biotecnologia, Genética e Biologia Celular, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

Laboratório Fenn de Espectrometria de Massas, Universidade Federal do Paraná (UFPR), 86900-000 Jandaia do Sul-PR, Brazil

Departamento de Química, Universidade Estadual de Maringá (UEM), 87020-900 Maringá-PR, Brazil

https://orcid.org/0000-0003-4835-7773

* e-mail: eduardo.meurer@ufpr.br

Editor handled this article: Eduardo Carasek

Experiment	Crude sample		Enzyme/Substrate		NaOH/Substrate
Experiment	Level	Concentration / %	Level	Concentration / %	Level	Concentration / %
A	+1	100	+1	10	+1	100
B	+1	100	−1	1	−1	50
C	−1	50	+1	10	+1	100
D	−1	50	−1	1	−1	50
CP	0	75	0	5.5	0	75

Experiment	Crude sample	Chemical/S	Pt / %
Experiment	Crude sample	Chemical/S	NaOH	Neutrase®	Alcalase®	Flavourzyme®
A	+1	+1	2.86 ± 0.1	5.97 ± 0.3	8.28 ± 0.4	3.80 ± 0.1
B	+1	−1	2.93 ± 0.1	4.50 ± 0.2	7.64 ± 0.2	3.25 ± 0.1
C	−1	+1	2.36 ± 0.1	3.52 ± 0.1	4.55 ± 0.5	1.82 ± 0.2
E	−1	−1	1.82 ± 0.2	2.89 ± 0.3	4.29 ± 0.2	1.28 ± 0.1
CP	0	0	2.60 ± 0.1	4.55 ± 0.6	6.58 ± 0.2	2.70 ± 0.11

Experiment	Crude sample	Chemical/S	DH / %
Experiment	Crude sample	Chemical/S	NaOH	Neutrase®	Alcalase®	Flavourzyme®
A	+1	+1	32.68 ± 2.4	24.41 ± 0.6	33.35 ± 0.1	49.48 ± 1.3
B	+1	−1	28.16 ± 4.2	27.88 ± 1.2	31.22 ± 0.3	45.35 ± 0.6
C	−1	+1	18.24 ± 1.1	19.54 ± 1.2	30.40 ± 0.7	61.80 ± 0.1
D	−1	−1	16.81 ± 2.2	21.31 ± 0.4	28.90 ± 0.2	59.83 ± 0.7
CP	0	0	30.51 ± 3.8	23.67 ± 0.6	34.78 ± 0.1	60.11 ± 1.7

Treatment	NaOH / %	Neutrase® / %	Alcalase® / %	Flavourzyme® / %
Crude sample	89	100	80	50
Chemical/S	100	1	6	10
Theoretical result	33.93	27.95	35.83	63.00
Experimental result	32.68 ± 2.4	27.88 ± 1.2	33.64 ± 1.6	61.80 ± 0.6

Treatment	NaOH / %	Neutrase® / %	Alcalase® / %	Flavourzyme® / %
Crude sample	100	100	100	100
Chemical/S	50	10	10	10
Theoretical result	2.95	6.03	8.28	3.83
Experimental result	2.93 ± 0.1	5.97 ± 0.3	8.28 ± 0.4	3.80 ± 0.1

Treatment		Optimal samples
Treatment		NaOH		Neutrase®		Alcalase®		Flavourzyme®
Amino acid	m/z	DH	Pt	DH	Pt	DH	Pt	DH	Pt
Serine	106	–	501^a	557^a	907^b	393^a	380^a	432^a	564^a
Proline	116	1319^a	1589^a	707^a	1806^a	2070^a	657^a	7551^a, ^b	17480^b
Valine	118	233^a	911^a, ^b, ^c	639^a, ^b	1958^d	1638^c, ^d	616^a, ^b	1324^b, ^c, ^d	1743^d
Isoleucine/Leucine	132	739^a	3890^a	1487^a	14437^b	10956^b	1947^a	11530^b	10942^b
Histidine	156	239^a	435^a, ^b, ^c	449^a, ^b, ^c	801^c	387^a, ^b	372^a, ^b	515^a, ^b, ^c	665^b, ^c
Fenilalanine	166	5578^a	7625^a, ^b	2060^a	23375^c	21921^b, ^c	5185^a	36007^c	27568^c
Tirosine	182	251^a	62^a, ^b	249^a	803^b	708^a, ^b	–	607^a, ^b	631^a, ^b

Ions	Peptides	Optimal samples
Ions		NaOH		Neutrase®		Alcalase®		Flavourzyme®
m/z		DH	Pt DH	Pt	DH	Pt	DH	Pt
177	GT	322^a	271^a	368^a	1.057^c	279^a	236^a	798^b, ^c	524^b
205	SV	761^a	1658^a	836^a	1181^a	1388^a	540^a	8999^b	7048^b
243	SH	112^a	273^a	293^a	1101^c	206^a	2088^d	709^b	1555^c
245	(I/L)(I/L)	238^a	–	152^a	1054^b	375^a	1790^b	–	286^a
277	VGT; SG(I/L)	180^a	167^a	126^a	704^a, ^b	143^a	1262^b	189^a	409^a
279	F(I/L); (I/L)F	708^a, ^b	442^a, ^b	292^a	1464^b, ^c	387^a, ^b	2240^c	242^a	328^b
309	GMT	453^a	618^a	393^a	593^a	242^a	614^a	299^a	548^a
313	FF	300^a	249^a	132^a	779^b	347^a	877^c	220^a	240^a

Ions							Sequence	Biological activity	Reference
[M + H]⁺	y1	Residue	Imonium	Related	b2	y2	Sequence	Biological activity	Reference
177	120(T)	57(G)	–	–	159	–	GT	ACE inhibitor	23
205	118(V)	87(S)	–	–	187	–	SV	DPPIV inhibitor	24
243	156(H)	87(S)	–	–	225	–	SH	DPPIV inhibitor	24
245	132(I/L)	113(I/L)	86	-	227	–	(I/L)(I/L)	DPPIV inhibitor; glucose stimulant and ACE inhibitor	24-27
277	120(T)	157(VG)	72	-	157	177	VGT	-	-
	132(I/L)	145(SG)	86	72	145	189	SG(I/L)	-	-
279	132(I/L)	147(F)	120	-	261	–	F(I/L)	DPPIII and DPPIV inhibitor	28,29
	166(F)	113(I/L)	120	-	261	–	(I/L)F	ACE inhibitor	23,30
309	120(T)	189(MG)	-	-	189	251	GMT	-	-
313	166(F)	147(F)	120	-	295	–	FF	ACE inhibitor; DPPIV inhibitor	31

[1] * e-mail: eduardo.meurer@ufpr.br

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Optimization of Hydrolysis and Identifcation of Bioactive Peptides in Brewery Yeast Residuals