title: Lactobacillus gasseri requires peptides, not proteins or free amino acids, for growth in milk.
abstract: Lactobacillus gasseri is a widespread commensal lactic acid bacterium inhabiting human mucosal niches and has many beneficial effects as a probiotic. However, L. gasseri is difficult to grow in milk, which hurts usability for the food industry. It had been previously reported that supplementation with yeast extract or proteose peptone, including peptides, enables L. gasseri to grow well in milk. In this study, our objective was to confirm peptide requirement of L. gasseri and evaluate efficacy of peptide release by enzymatic proteolysis on growth of L. gassei in milk. Three strains of L. gasseri did not grow well in modified DeMan, Rogosa, Sharpe broth without any nitrogen sources (MRS-N), but addition of a casein-derived peptide mixture, tryptone, promoted growth. In contrast, little effect was observed after adding casein or a casein-derived amino acid mixture, casamino acids. These results indicate that L. gasseri requires peptides, not proteins or free amino acids, among milk-derived nitrogen sources for growth. Lactobacillus gasseri JCM 1131T hardly had growth capacity in 6 kinds of milk-based media: bovine milk, human milk, skim milk, cheese whey, modified MRS-N (MRSL-N) supplemented with acid whey, and MRSL-N supplemented with casein. Moreover, treatment with digestive proteases, particularly pepsin, to release peptides made it grow well in each milk-based medium. The pepsin treatment was the most effective for growth of strain JCM 1131T in skim milk among the tested food-grade proteases such as trypsin, α-chymotrypsin, calf rennet, ficin, bromelain, and papain. As well as strain JCM 1131T, pepsinolysis of milk improved growth of other L. gasseri strains and some strains of enteric lactobacilli such as Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus johnsonii, and Lactobacillus reuteri. These results suggest that some relatives of L. gasseri also use peptides as desirable nitrogen sources, and that milk may be a good supplier of nutritious peptides to enteric lactobacilli including L. gasseri after peptic digestion in the gastrointestinal tract. This is the first report showing peptide requirement of L. gasseri and efficacy of pepsinolysis on the growth of L. gasseri and its relatives in milk. This study would contribute to increasing usability of L. gasseri and its relatives as probiotics in dairy foods.
authors.affiliation: Graduate School of Environmental and Life Science, Okayama University, Okayama 7008530, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Arakawa
authors.affiliation: Graduate School of Environmental and Life Science, Okayama University, Okayama 7008530, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Matsunaga
authors.affiliation: Graduate School of Environmental and Life Science, Okayama University, Okayama 7008530, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Takihiro
authors.affiliation: Graduate School of Environmental and Life Science, Okayama University, Okayama 7008530, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Moritoki
authors.affiliation: Graduate School of Environmental and Life Science, Okayama University, Okayama 7008530, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Ryuto
authors.affiliation: Graduate School of Bioresource Sciences, Nihon University, Kanagawa 2520880, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kawai
authors.affiliation: Graduate School of Bioresource Sciences, Nihon University, Kanagawa 2520880, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Masuda
authors.affiliation: Graduate School of Environmental and Life Science, Okayama University, Okayama 7008530, Japan. Electronic address: tmnono@cc.okayama-u.ac.jp.
authors.forename: T
authors.initials: T
authors.lastname: Miyamoto
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002364:Caseins / Q000378:metabolism; D002417:Cattle; D002918:Chymotrypsin / Q000378:metabolism; D041981:Gastrointestinal Tract / Q000382:microbiology; D007778:Lactobacillus / Q000254:growth & development*; D008892:Milk / Q000737:chemistry / Q000382:microbiology*; D009584:Nitrogen / Q000737:chemistry; D010434:Pepsin A / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D019936:Probiotics / Q000378:metabolism
chemical_list: D000596:Amino Acids; D002364:Caseins; D010446:Peptide Fragments; D010455:Peptides; C017721:casamino acids; C018135:proteose-peptone; D010447:Peptide Hydrolases; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D010434:Pepsin A; D009584:Nitrogen
keywords: Lactobacillus gasseri; pepsin; peptide requirement; proteolysis