Purification and Characterization of a Novel Serine Aminopeptidase from <i>Lactobacillus casei</i> Ssp. <i>casei</i> IFPL 731

Fernández-Esplá, M. D.; Fox, Patrick F.; Martín-Hernández, M. C.

doi:10.1021/jf960889w

Cited by 9 publications

(2 citation statements)

References 26 publications

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“…1997a,b; Fernández‐Esplá & Martín‐Hernández. 1997; Fernández‐Esplá et al . 1997a,b), makes this micro‐organism a valuable adjunct to cheese starters, providing the medium, on autolysis of the cells, with a collection of enzymes capable of enhancing cheese proteolysis and lipolysis.…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of an intracellular esterase fromLactobacillus caseisubsp.caseiIFPL731

Castillo

Requena

Palencia

et al. 1999

Journal of Applied Microbiology

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An intracellular esterase from Lactobacillus casei subsp. casei IFPL731 was purified 1000‐fold by ion exchange chromatography and gel filtration chromatography. The relative molecular mass of the native enzyme was 105 kDa, while the subunit molecular mass was estimated to be 38 kDa. The esterase hydrolysed tributyrin and had a preference for esters of short‐chain fatty acids (butyrate, caproate and caprylate), while it did not hydrolyse palmitate and sterate esters. The apparent Michaelis‐Menten constant of the enzyme on p‐nitrophenyl butyrate was 0·3 mmol l−1 while on p‐nitrophenyl caprylate, it was 0·04 mmol l−1. The esterase was active over a broad range of pH and temperature values, and retained about 50% of maximal activity at pH 5·0 and 12 °C. Activity was strongly inhibited by 5 mmol l−1 phenylmethylsulphonyl fluoride, β‐mercaptoethanol and N‐ethylmaleimide, and was stimulated by Zn2+ at 1 mmol l−1.

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Section: Discussionmentioning

confidence: 99%

Isolation and characterization of an intracellular esterase fromLactobacillus caseisubsp.caseiIFPL731

Castillo

Requena

Palencia

et al. 1999

Journal of Applied Microbiology

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“…Beads, cell debris and unbroken cells were removed by centrifugation (10,000 × g at 4°C for 5 min) and the supernatant fluid was designated as the intracellular enzymatic extract (IE) which was maintained on ice and immediately used for enzymatic assays. Intracellular aminopeptidase activity was assayed with the chromogenic substrate Lys-ρNa (Sigma) as previously described (29) by incubation of 100 μl of IE with 400 μl of Tris-HCl buffer (50 mM, pH 7.0) and 50 μl Lys-ρNa (10 mM) at five temperatures (35, 40, 45, 50, and 55°C) for 20 min to determine the effect of temperature on the aminopeptidase activity. The reaction was stopped by addition of 1 mL of 30% acetic acid.…”

Section: Methodsmentioning

confidence: 99%

Use of Mass Spectrometry to Profile Peptides in Whey Protein Isolate Medium Fermented by Lactobacillus helveticus LH-2 and Lactobacillus acidophilus La-5

et al. 2019

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Peptides in the 3-kDa ultrafiltrate of fermented whey protein isolate (WPI) medium could be responsible for the antivirulence activity of Lactobacillus helveticus LH-2 and Lactobacillus acidophilus La-5 against Salmonella Typhimurium. Non-fermented and fermented media containing 5.6% WPI were fractionated at a 3 kDa cut-off and the filtrate was analyzed by mass spectrometry. The non-fermented WPI medium contained 109 milk derived peptides, which originated from β-casein (52), αs1-casein (22), αs2-casein (10), κ-casein (8), and β-lactoglobulin (17). Most of these peptides were not found in the fermented media, except for 14 peptides from β-casein and one peptide from αs2-casein. Database searches confirmed that 39 out of the 109 peptides had established physiological functions, including angiotensin-converting-enzyme (ACE) inhibitory, antioxidant, antimicrobial, or immunomodulating activity. A total of 75 peptides were found in the LH-2 cell free spent medium (CFSM): 54 from β-casein, 14 from k-casein, 4 from β-lactoglobulin and 3 from αs2-casein. From these peptides, 19 have previously been associated with several categories of bioactivity. For La-5 CFSM, a total of 15 peptides were sequenced: 8 from β-casein, 5 from αs1-casein, 2 from β-lactoglobulin. Only 5 of these have previously been reported as having bioactivity. Many of the peptides remaining in the fermented medium would contain low-affinity residues for oligopeptide binding proteins and higher resistance to peptidase hydrolysis. These properties of the sequenced peptides could explain their accumulation after fermentation despite the active proteolytic enzymes of LH-2 and La-5 strains. Down-regulated expression of hilA and ssrB genes in S. Typhimurium was observed in the presence of La-5 and LH-2 CFSM. Downregulation was not observed for the Salmonella oppA mutant strain exposed to the same CFSM used to treat the S. Typhimurium DT104 wild-type strain. This result suggests the importance of peptide transport by S. Typhimurium for down regulation of virulence genes in Salmonella.

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Biochemistry of Cheese Ripening: Proteolysis

Ardö

McSweeney

Magboul

et al. 2017

Cheese

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Purification and Characterization of a Novel Serine Aminopeptidase from Lactobacillus casei Ssp. casei IFPL 731

Cited by 9 publications

References 26 publications

Isolation and characterization of an intracellular esterase fromLactobacillus caseisubsp.caseiIFPL731

Isolation and characterization of an intracellular esterase fromLactobacillus caseisubsp.caseiIFPL731

Use of Mass Spectrometry to Profile Peptides in Whey Protein Isolate Medium Fermented by Lactobacillus helveticus LH-2 and Lactobacillus acidophilus La-5

Biochemistry of Cheese Ripening: Proteolysis

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