Overproduction of Serratia marcescens metalloprotease (SMP) from the recombinant Serratia marcescens strains

Kim, Ki-Seok; Park, Koon Sig; Byun, Sun‐Sig; Pan, Jae-Gu; Shin, Yong Chul

doi:10.1007/bf00132017

Cited by 8 publications

(2 citation statements)

References 11 publications

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“…The amino acids in the medium are due to the breakdown of protein (TF) that eventually increased as the time increased indicating the degradation of the protein. Enhanced protease production by fish gut microflora utilizing TF, a protein‐rich source containing all amino acids, is in agreement with the results of previous researchers using protein‐rich media such as casein, peptone, tryptone, gelatin, skim milk, amino acid leucine , and different Bacillus strains, B. subtilis A26 (269.36 U/mL), Bacillus sp. L21 (306.5 U/mL), Bacillus sp.…”

Section: Resultssupporting

confidence: 71%

Production of different proteases from fish gut microflora utilizing tannery fleshing

Sumathi

Mohanapriya

Mandal

et al. 2012

Engineering in Life Sciences

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Three alkaline protease-producing strains designated as ANFLR1, NPLR1, and PROLR15 were isolated from Labeo rohita fish gut. These strains are able to produce alkaline protease using tannery fleshing (TF) as the sole carbon and nitrogen source and were identified as Bacillus megaterium, Serratia marcescens, and novel Pontibacter sps. Proteases from these organisms were purified to electrophoretic homogeneity following ammonium sulphate precipitation, ion exchange, and column chromatography. SDS-PAGE revealed molecular weights of the proteases to be 46 kDa (ANFLR1), 52 kDa (NPLR1), and 58 kDa (PROLR15). The optimum pH and temperature for the protease activity of ANFLR1, NPLR1, and PROLR15 were found to be 10. 5, 11.5, 9, and 70 • C, 60 • C, and 50 • C, respectively. The maximum protease activities at the optimum conditions were 420 U/mL (ANFLR1), 550 U/mL (NPLR1), and 530 U/mL (PROLR15). Inhibition of the NPLR1 protease by pepstatin confirmed aspartate-type enzymatic activity. Fe 3+ enhanced the activity of PROLR15 protease. Unlike all other microbial proteases known so far, the PROLR15 enzyme did not require Ca 2+ for activity and thermal stability. SDS-PAGE and scanning electron microscopy analyses confirmed the conversion of high molecular weight substrate (TF) to low molecular weight peptides by these proteases. The alkaline metalloprotease production by novel Pontibacter sps. and aspartate protease production by S. marcescens remain unexplored. Hence, TF with its relatively abundant availability can be beneficially utilized for alkaline protease production through the fish gut microbial fermentation processes.C. Sumathi et al. Eng. Life Sci. 2012, 12, No. 2, 223-237 proteases [3,9,[12][13][14][15]. The strains identified are featherdegrading Bacillus licheniformis [16-18], Bacillus subtilis [19], Bacillus cereus [20], Bacillus pseudofirmus [21] keratinolytic proteases that are useful in dehairing of animal hides, B. subtilis [22-24], Bacillus velesensis [25], B. cereus [26], B. licheniformis [27], and Bacillus pumilus [28], which subsequently yield the edible protein hydrolysates from fish, lean meat, Zein, fresh whey [12], and shrimp shell wastes [29,30] through fermentation processes [31][32][33]. Isolation of three different alkaline proteaseproducing bacterial strains ANFLR1, NPLR1, and PROLR15 after adaptation with TF is a novel approach which has not been explored yet. Alkaline proteases generally belong to the group of proteases, which have either a serine center or a metallo-type, exhibiting a wide pH range (pH 6-13). Similarly, Gram-negative bacilli Serratia sp. cultivated in proteinaceous substrates generally produce two different kinds of extracellular proteases-a major metalloprotease and a minor serine protease [10,34]. This is the first approach to reveal aspartate alkaline protease production by a Serratia strain. Furthermore, the present work explores the unexploited application of alkaline protease production by a novel Pontibacter strain PROLR15 isolated from the novel source Labeo roh...

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Section: Resultssupporting

confidence: 71%

Production of different proteases from fish gut microflora utilizing tannery fleshing

Sumathi

Mohanapriya

Mandal

et al. 2012

Engineering in Life Sciences

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“…S. aureus produce different types of proteases widely studied and classified into three types; serine, thiol and metallo proteases [13]. Nowadays the economic importance of metallo proteases in some application, especially as an anti-inflammatory agent is well known [14].…”

Section: Introductionmentioning

confidence: 99%

Effect of aeration and agitation on the protease production by Staphylococcus aureus mutant RC128 in a stirred tank bioreactor

Ducrós

Ferrari

Pellegrino

et al. 2008

Bioprocess Biosyst Eng

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The modified rotating simplex method has been successfully used to determine the best combination of agitation rate and aeration rate for maximum production of extracellular proteases by Staphylococcus aureus mutant RC128, in a stirred tank bioreactor operated in a discontinuous way. This mutant has shown altered exoprotein production, specially enhanced protease production. Maximum production of proteases (15.28 UP/ml), measured using azocasein as a substrate, was obtained at exponential growth phase when the bioreactor was operated at 300 rpm and at 2 vvm with a volumetric oxygen transfer coefficient (K(L)a) of 175.75 h(-1). These conditions were found to be more suitable for protease production.

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Purification, characterization, and properties of an alkaline protease produced bySerratia marcescensS3‐R1inhabiting Korean ginseng rhizosphere

et al. 2013

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Overproduction of Serratia marcescens metalloprotease (SMP) from the recombinant Serratia marcescens strains

Cited by 8 publications

References 11 publications

Production of different proteases from fish gut microflora utilizing tannery fleshing

Production of different proteases from fish gut microflora utilizing tannery fleshing

Effect of aeration and agitation on the protease production by Staphylococcus aureus mutant RC128 in a stirred tank bioreactor

Purification, characterization, and properties of an alkaline protease produced bySerratia marcescensS3‐R1inhabiting Korean ginseng rhizosphere

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