Characterization of Trypsin-Like Protease of Lactobacillus plantarum FNCC 0270

Abstract

Trypsin is an enzyme that has a unique mechanism of cutting peptide bonds specifically at the carboxyl side of lysine or arginine amino acids, with another amino acid. This study aims to analyze a trypsin-like protease (TLP) found in Lactobacillus plantarum FNCC 0270, by performing partial proteomic tests, i.e. MALDI-TOF/TOF, and standard bioinformatics tools. SDS-PAGE analysis showed 4 protein bands. Two bands of the (P 1 and P 2 ) showed molecular weights equivalent to 47.35 and 38.42 kD, each generating 8 and 11 peptide fragments respectively. According to information in www.ncbi.nlm.nih.gov/genbank/structures, the structure of serine protease HtrA (subs. plantarum L. plantarum ST-III) consists of three domains. Using Clone Manager ® software by aligning two sequences we obtained eleven. The Lactobacillus produces of the trypsin-like serine protease has 40-90% similarity. Using the Clustal W2 software we passed the 11 sequences through multiple alignments, and found that the isolate L. plantarum is closely related to L. buchneri, L. brevis, and L. malefermentans on the phylogenetic tree. Alignment analysis results showed that all 8 peptide fragments of band 1 and 11 peptide fragments of band 2, of the SDS-PAGE, were located in the active domain region of the fourth trypsin-like serine protease producing Lactobacilli.

Effect of Lactobacillus plantarum or Enterococcus faecalis as co‐inoculants with Aspergillus oryzae in koji making on the physicochemical properties of soy sauce

“…Compared with the control group, neutral protease activity of the experimental group ( A. oryzae + NSTLAB‐LE) was slightly decreased (Figure 1b), while acid protease activity of group A ( L. plantarum + A. oryzae ) and group B ( E. faecalis + A. oryzae ) increased by 50.9% and 54.6%, respectively ( p < 0.05) at 36 h of koji making (Figure 1a), which can be attributed to incorporation of NSTLAB‐LE to koji making. Previous studies have described that these two bacterial species can produce acid protease (Margono et al., 2014; Ramakrishnan et al., 2012), and high protease activity promotes protein decomposition and production of certain amino acids and oligopeptides that are beneficial to the growth of NSTLAB‐LE (Hebert et al., 2004).…”

Wide-genome selection of lactic acid bacteria harboring genes that promote the elimination of antinutritional factors

“…Although several trypsins and trypsin-like proteases (TLPs) may be manufactured by microorganisms such as Trichoderma and Bacillus , this may have a number of risk for use in therapeutic drugs. TLP enzymes can execute the same function as trypsin and can be synthesized by LAB (e.g., Wulansari et al., 2012 ; Margono et al., 2014 ). In this study, 15 out of 150 genomes of Enterobacteriaceae have only one (in total three different genes) gene encoding for trypsin; 36 out of 155 genomes of Lactobacillaceae harbor one gene are involved in the synthesis of trypsin-like serine protease with two different genes ( Supplementary Table S2 ).…”

Optimization of flaxseed milk fermentation for the production of functional peptides and estimation of their bioactivities

“…The remarkable proteolytic activity of the probiotic strains is possibly due to their ability to release extracellular and intracellular proteases during fermentation (Fang et al, 2018;Indarmawan et al, 2016). Enzymes such as serine protease HtrA, dipeptidase, prolyl endopeptidyl peptidase, prolinase, prolidase, and iminopeptidase have been linked to Lactobacilli spp, especially L. plantarum (Francavilla et al, 2017;Margono et al, 2014).…”