Bioconversion of squid pen by Lactobacillus paracasei subsp paracasei TKU010 for the production of proteases and lettuce growth enhancing biofertilizers.
A protease-producing bacterium, strain TKU010, was isolated from infant vomited milk and identified as Lactobacillus paracasei subsp. paracasei. A surfactant-stable protease, purified 64-fold from the third day culture supernatant to homogeneity in an overall yield of 11%, has a molecular weight of about 49,000. The enzyme degraded casein and gelatin, but did not degrade albumin, fibrin, and elastin. The enzyme activity was increased about 1.5-fold by the addition of 5mM Ba(2+). However, Fe(2+) and Cu(2+) ions strongly inhibited the enzyme. The enzyme was maximally active at pH 10 and 60 degrees C and retained 94% and 71% activity in the presence of Tween 20 (2% w/v) and SDS (2mM), respectively. The result of identification of TKU010 protease showed that nine tryptic peptides were identical to Serratia protease (serralysin) (GenBank accession number gi999638) with 35% sequence coverage. In comparison with the tryptic peptides of L. paracasei subsp. paracasei TKU012 protease, TKU010 protease possessed two additional peptides with sequences of AATTGYDAVDDLLHYHER and QTFTHEIGHALGLSHPGDYNAGEGNPTYR. The fourth day culture supernatant of TKU010 showed maximal activity of about 5-fold growth enhancing effect on lettuce weight, which was not shown with L. paracasei subsp paracasei TKU012.
Wang
San-Lang
SL
Graduate Institute of Life Sciences, Tamkang University, Taipei 251, Taiwan. sabulo@mail.tku.edu.tw
Huang
Tsai-Yi
TY
Wang
Chun-Yuan
CY
Liang
Tzu-Wen
TW
Yen
Yue-Horng
YH
Sakata
Yusuke
Y
D000595:Amino Acid Sequence; D000818:Animals; D001937:Brassica / Q000254:growth & development; D002413:Cations, Divalent / Q000378:metabolism; D049832:Decapodiformes; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007778:Lactobacillus / Q000201:enzymology* / Q000254:growth & development; D018545:Lactuca / Q000254:growth & development*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D012705:Serratia / Q000201:enzymology; D013379:Substrate Specificity; D013501:Surface-Active Agents / Q000378:metabolism
D002413:Cations, Divalent; D010446:Peptide Fragments; D013501:Surface-Active Agents; D010447:Peptide Hydrolases