Repressive processing of antihypertensive peptides, Val-Pro-Pro and Ile-Pro-Pro, in Lactobacillus helveticus fermented milk by added peptides

Wakai, Taketo; Yamaguchi, Naoya; Hatanaka, Minoru; Nakamura, Yasunori; Yamamoto, Naoyuki

doi:10.1016/j.jbiosc.2012.03.015

Cited by 18 publications

(25 citation statements)

References 32 publications

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“…To evaluate the binding of the BCARR protein to specific DNA fragments containing the promoters of the pepO, pepO2, pepT2, pepCE and dppD genes, which are negatively regulated in response to added peptides [11], EMSAs were carried out in the presence of BCAAs. The results demonstrated that DNA fragments from upstream of the pepO , pepO2 , pepT2 , pepCE and dppD genes were shifted when the BCARR protein and BCAAs were present in the reaction mixture (Figure 4A).…”

Section: Resultsmentioning

confidence: 99%

“…Six pairs of primers were designed to amplify the promoter regions (approximately 300 bp each) controlling expression of 6 proteolysis genes (Table 1) known to be down-regulated in the presence of peptides in L. helveticus CM4 [11]. To amplify the promoters for the pepCE and pepO genes, which are not the first genes in each operon, the putative promoter region of the first gene in each operon was used.…”

Section: Methodsmentioning

confidence: 99%

“…Primers for the pepV gene (pepVRTF, pepVRTR) and the internal reference, glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) gene (gapRTF, gapRTR) were used. To understand repressed gene expression of the proteolytic system in milk medium [11], E. coli transformants were cultured in M9 medium [20] with or without both 0.4% casamino acids (hydrolysate of milk casein: BD) and 10 mM BCAAs. The cells were cultured in M9 and M9+ BCAAs medium and harvested at optical densities (OD 600 ) of 0.6 and 0.9, one hour after each inoculation, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…In the previous study, the existence of a CodY-like regulatory system controlling expression of the pepO , pepO2 , pepV , pepCE , pepT2 and dppD genes in response to amino acids was suggested by transcriptome analysis of L. helveticus CM4 [11]. However, there are no CodY-like homologs in lactobacilli and no reports of a regulator in lactobacilli to explain the repressive effect on the proteolytic system by amino acids.…”

Section: Introductionmentioning

confidence: 96%

“…A highly active cell wall-associated proteinase and several intracellular peptidases specific to L. helveticus strain CM4 are thought to be responsible for the organism’s ability to release large amounts of these two antihypertensive tripeptides [9]. However, the production of VPP and IPP by L. helveticus CM4 is mildly repressed by peptides [10] and amino acids that accumulate in fermented milk as a result of the down-regulation of genes such as pepO2 , pepCE and pepE , which most likely encode enzymes involved in the processing of both bioactive peptides [11]. In Lactococcus lactis , most of the genes regulated by the CodY protein in response to branched chain amino acids (BCAAs) are involved in the proteolysis system [12].…”

Section: Introductionmentioning

confidence: 99%

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A Novel Branched Chain Amino Acids Responsive Transcriptional Regulator, BCARR, Negatively Acts on the Proteolytic System in Lactobacillus helveticus

Wakai¹,

Yamamoto²

2013

PLoS ONE

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Transcriptional negative regulation of the proteolytic system of Lactobacillus helveticus CM4 in response to amino acids seems to be very important for the control of antihypertensive peptide production; however, it remains poorly understood. A 26-kDa protein with N-terminal cystathionine β-synthase domains (CBS domain protein), which seems to be involved in the regulatory system, was purified by using a DNA-sepharose bound 300-bp DNA fragment corresponding to the upstream regions of the six proteolytic genes that are down-regulated by amino acids. The CBS domain protein bound to a DNA fragment corresponding to the region upstream of the pepV gene in response to branched chain amino acids (BCAAs). The expression of the pepV gene in Escherichia coli grown in BCAA-enriched medium was repressed when the CBS domain protein was co-expressed. These results reveal that the CBS domain protein acts as a novel type of BCAA-responsive transcriptional regulator (BCARR) in L. helveticus. From comparative analysis of the promoter regions of the six proteolysis genes, a palindromic AT-rich motif, 5′-AAAAANNCTWTTATT-3′, was predicted as the consensus DNA motif for the BCARR protein binding. Footprint analysis using the pepV promotor region and gel shift analyses with the corresponding short DNA fragments strongly suggested that the BCARR protein binds adjacent to the pepV promoter region and affects the transcription level of the pepV gene in the presence of BCAAs. Homology search analysis of the C-terminal region of the BCARR protein suggested the existence of a unique βαββαβ fold structure that has been reported in a variety of ACT (aspartate kinase-chorismate mutase-tyrA) domain proteins for sensing amino acids. These results also suggest that the sensing of BCAAs by the ACT domain might promote the binding of the BCARR to DNA sequences upstream of proteolysis genes, which affects the gene expression of the proteolytic system in L. helveticus.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

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A Novel Branched Chain Amino Acids Responsive Transcriptional Regulator, BCARR, Negatively Acts on the Proteolytic System in Lactobacillus helveticus

Wakai¹,

Yamamoto²

2013

PLoS ONE

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Production of an antihypertensive peptide from milk by the brown rot fungus Neolentinus lepideus

Okamoto

Kawamura

Tagawa

et al. 2020

Eur Food Res Technol

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Functional peptides in Asian protein rich fermented foods: production and health benefits

Kumari

Sanjukta

Kumar

2021

Syst Microbiol and Biomanuf

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The richness and diversity of protein rich fermented foods are chiefly observed in many Asian countries. They are potential source of wide range of biologically active peptides depending on the protein source and starter culture used for fermentation. The major protein rich fermented foods popularly consumed in Asian countries include fermented soybean, fermented milk and fermented fish products. These foods are fermented using different starter cultures including lactic acid bacteria (LAB), Bacillus species, yeast and filamentous fungi. Fermentation by individual starter culture results in unique products by a series of enzymatic hydrolysis, leading to production of a diversity of bioactive peptides responsible for health benefits. Bioactive peptides in Asian protein rich fermented foods have been reported for exhibiting antihypertensive, antioxidant, antibacterial, antidiabetic and anticancer properties. This review is focused on Asian protein rich fermented foods, associated microbes and bioactive peptides responsible for health benefits.

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Repressive processing of antihypertensive peptides, Val-Pro-Pro and Ile-Pro-Pro, in Lactobacillus helveticus fermented milk by added peptides

Cited by 18 publications

References 32 publications

A Novel Branched Chain Amino Acids Responsive Transcriptional Regulator, BCARR, Negatively Acts on the Proteolytic System in Lactobacillus helveticus

A Novel Branched Chain Amino Acids Responsive Transcriptional Regulator, BCARR, Negatively Acts on the Proteolytic System in Lactobacillus helveticus

Production of an antihypertensive peptide from milk by the brown rot fungus Neolentinus lepideus

Functional peptides in Asian protein rich fermented foods: production and health benefits

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