• Journal of Microbiology and Biotechnology
• /
• 제2권4호
• /
• pp.231-236
• /
• 1992

A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular serine protease gene was cloned in Escherichia coli. The cloned gene(pSMP18) caused specific excretion of the protease into the extracellular medium through the outer membrane of E. coli host cells. The protease purified from E. coli harboring pSMP18 was inactivated not by 100 mM EDTA but by 10 mM phenyl methyl sulfonyl flouride (PMSF). The molecular weight of the purified serine protease was about 66, 000 in the SDS-PAGE and the isoelectric point was approximately 5.7 in IEFㆍGel electrophoresis. The optimal pH and temperature for reaction of the purified serine protease were 9.5 and $45^\circ{C}$, respectively.

• Journal of Microbiology and Biotechnology
• /
• 제17권10호
• /
• pp.1622-1628
• /
• 2007

We characterized the efficacy of an electrostatic precipitator (ESP) air-cleaner in reducing the concentration of Serratia marcescens in an enclosed space. We used an experimental room ($4.5{\times}3{\times}2.9\;m$) in which electrostatic air-cleaners were located. Two air-cleaners enhanced the equivalent ventilation rates in the chamber by about 3.3 air changes per hour (ACH) over the 2 ACH provided by the mechanical ventilation system. Natural die-off of the organisms provided an additional equivalent of 3 ACH, so that the total ventilation rate with the ESP air-ccleaners was 8.3 ACH. We also examined whether the ESP air-cleaners altered the deposition of Serratia marcescens aerosols on the experimental room surfaces. We did not find any significant differences in the number of colony forming units recovered from surfaces with and without the air-cleaners. We installed UV lights inside the ESPs and determined if UV light, in addition to electrical fields, increased the efficacy of the ESPs. The presence of UV light inside the ESP reduced S. marcescens aerosols by approximately 2 ACH. Finally, a box model indicates that the efficiency of the air-cleaner increases for both biological and nonbiological particles at ventilation rates of 0.2-1, which are typical for residential settings.

• 미생물학회지
• /
• 제30권6호
• /
• pp.507-513
• /
• 1992

Serratia marecescens RH1 isolated from soil samples produced large amount of extracellular proteases. One of the genes encoding an extracellular protease form S. marcescens RH1 was cloned in Escherichia coli by shot gun cloning method. The cloned protease, SSP, was stably expressed by its own promoter and excreted into the extracellular medium from E. coli host (ORF) of 3.135 nucleotides corresponding to 1.045 amino acids (112 kDa). The nucleotide and deduced amino acid sequence of SSP showed high overall homology (88%) to one of the S. marcescens protease (27), but low homology to other serine protease families. The optimal pH and temperature of the enzyme were pH 9.0 and 45.deg.C respectively. The activity of protease was inhibited by phenylmethylsulfonyl fluoride (PMSF), which suggests that the enzyme is a serine protease.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제3권2호
• /
• pp.71-77
• /
• 1998

The strain of Serratia marcescens QM B1466 produces selectively large amount of chitinolytic enzymes (about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acetyl-${\beta}$-D-glucosamine (NAG) was performed with a system consisting of two hydrolases (chitinase and chitobiase) produced by optimization of a microbial host consuming chitin particles. For the development of Large-scale biological process for the production of NAG from chitinaceous waste, the selection and optimization of a microbial host, particle size of crab/shrimp chitin sources and initial induction time using chitin as a sole carbon source on chitinase/chitobiase production and NAG production were examined. Crab-shell chitin(1.5%) treated by dilute acid and , ball-milled with a normal diameter less than 250m gave the highest chitinase activity over a 7 days culture. Crude chitinase/ chitobiase solution obtained in a 10 L fed-batch fermentation showed a maximum activities of 23.6 U/mL and 5.1 U/mL, respectively with a feeding time of 3 hrs, near pH 8.5 at 30$^{\circ}C$.

• BMB Reports
• /
• 제28권1호
• /
• pp.40-45
• /
• 1995

Acetolactate synthase purified from Serratia marcescens ATCC 25419 was rapidly inactivated by the thiol specific reagent p-chloromercuribenzoate (PCMB), the tryptophan specific reagent N-bromosuccinimide (NBS), and the arginine modifying reagent phenylglyoxal (PGO). Inactivation by PCMB was prevented by both ${\alpha}$-ketobutyrate and pyruvate, and the second order rate constant for the inactivation was $2480\;M^{-1}{\cdot}min^{-1}$. The reaction order with respect to PCMB was 0.94. The inactivation of the enzyme by NBS was also substantially reduced by both ${\alpha}$-ketobutyrate and pyruvate. The second order rate constant for inactivation by NBS was $15,000\;M^{-1}{\cdot}min^{-1}$, and the reaction order was 2.0. On the other hand, inactivation by PGO was partially prevented by ${\alpha}$-ketobutyrate, but not by pyruvate. The second order rate constant for the inactivation was $1480\;M^{-1}{\cdot}min^{-1}$ and the order of reaction with respect to PGO was 0.75. These results suggest that essential cysteine, tryptophan and arginine are located at or near the substrate binding site.

• Archives of Plastic Surgery
• /
• 제41권4호
• /
• pp.414-417
• /
• 2014

Serratia marcescens (S. marcescens) emerged as an opportunist in the setting of immunodeficiency in the 1970s, when serious infections occurred in San Francisco hospitals after USA. Navy experiments had aerosolized the bacteria to study biologic warfare. We investigate the risks of S. marcescens in San Franciscans who undergo mastectomy with implant reconstruction. From 2007 to 2011, the senior author took breast capsule cultures for all patients at the time of tissue expander exchange/explant. Of the 142 women who had reconstruction, 23 had positive cultures. Only the two patients who were positive for S. marcescens developed clinical infections that required explantation. Both had postoperative chemotherapy with transient neutropenia, and both had close ties to San Francisco. Clinical signs of infection emerged for both patients months after initial surgery, despite having previously well healed incisions. Other patients were culture positive for Pseudomonas, Proteus, Enterococcus and MRSA and did not develop require explant. While the link between San Francisco and S. marcescens is controversial, a patient's geography is a simple screening tool when considering postoperative risks, especially in the immunocompromised. Closer monitoring for neutropenia during chemotherapy, and a lower threshold to administer S. marcescens targeted antibiotics may be warranted in these patients.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제7권1호
• /
• pp.38-42
• /
• 2002

A chitinase gene (pCHi58) encoding a 58 kDa chitinase was isolated from the Serratia marcescens KCTC 2172 cosmid library. The chitinase gene consisted of a 1686 bp open reading frame that encoded 562 amino acids. Escherichia coil harboring the pChi58 gene secreted a 58 kDa chitinase into the culture supernatant. The 58 kDa chitinase was purified using a chitin affinity column and mono-S column. A nucleotide and N-terminal amino acid sequence analysis showed that the 58 kDa chitinase had a leader peptide consisting of 23 amino acids which was cleaved prior to the 24th alanine. The 58 KDa chitinase exhibited a $98\%$ similarity to that of S. marcescens OMB 1466 in its nuclotide sequence. The chitinolytic patterns of the 58 kDa chitinase released N,N'-diacetyl chitobiose (NAG2) as the major hydrolysis end-product with a trace amount of N-acetylglucosamine. When a 4-methylumbellyferyl-N-acetylglucosamin monomer, dimmer, and tetramer were used as substrates, the 58 kDa chitinase did not digest the 4-Mu-NAG monomer $(analogue\;of\;NAG_2)$, thereby indicating that the 58 kDa chitinase was likely an endochitinase. The optimum reaction temperature and pH of the enzyme were $50^{\circ}C$ and 5.0, respectively.

• Journal of Microbiology and Biotechnology
• /
• 제20권5호
• /
• pp.881-888
• /
• 2010

solvent-tolerant bacterium strain, MH6, was isolated by hydrophilic organic solvent DMSO enrichment in the medium and identified as Serratia marcescens. The extracellular protease with novel organic-solvent-stable properties from strain MH6 was purified and characterized. The molecular mass of the purified protease was estimated to be 52 kDa on SDS-PAGE. The open reading frame (ORF) of the MH6 protease encoded 504 amino acids with 471 amino acid residues in the mature protease. Based on the inhibitory effects of EDTA and 1,10-phenathroline, the MH6 protease was characterized as a metalloproteinase. The enzyme activity was increased in the presence of $Ni^{2+}$, $Mg^{2+}$, and $Ca^{2+}$. The protease could also be activated by the nonionic surfactants Tween 80 (1.0%) and Triton X-100 (1.0%). The protease showed remarkable solvent stability in the presence of 50% (v/v) solutions of long-chain alkanes and long-chain alcohols. It was also fairly stable in the presence of 25% solutions of hydrophilic organic solvents. Owing to its high stability in solvents and surfactants, the MH6 protease is an ideal candidate for applications in organic catalysis and other related fields.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제28권2호
• /
• pp.25-31
• /
• 2014

Protaetia brevitarsis seulensis (Kolbe) is widely used in Korea, as a protein-rich, alternate, functional food with pharmacological benefits. In addition to anti-oxidant properties, the larvae of P. b. seulensis also show positive effects against hepatic disorder and diabetes; therefore, P. b. seulensis larvae are being reared on a large scale in Korea. We evaluated reared larvae of P. b. seulensis from Gyeong-gi in Korea. Using 16SrRNA PCR, electro-microscopy, and bioassay techniques, we found that the larvae harbored Spo-1, a bacterium identified as the insect pathogen Serratia marcescens. Therefore, we highlight the use of this insect as an alternate food and the need for its sanitary rearing conditions, as contamination may affect public health.

• Journal of Microbiology and Biotechnology
• /
• 제10권5호
• /
• pp.670-676
• /
• 2000

Several clones obtained from Serratia marcescens stimulated E. coli TP2139 (${\Delta}lac, \;{\Delta} crp$) cells to use maltose as a carbon source. The crp gene clone, pCKB12, was confirmed to stimulate the $\beta$-galactosidase activity, by Southern hybridization [31]. The nucleotide sequence of the crp region consisting of 1,979 bp was determined. The sequencing of the fragment led to the identification of two open reading frames: One of these, the crp gene, encoded 210 amino acid and the other encoded a truncated protein. The S. marcescens and E. coli crp genes showed a higher degree of divergence in their nucleotide sequence with 120 changes, however, the corresponding amino acid sequences showed only two amino acid differences. Yet, an analysis of the amino acid divergence revealed that the catabolite gene activator protein, the crp gene product, was the most conserved protein observed so far. Using a crp-lac protein fusion, it was demonstrated that S. marcescens CRP could repress its own expression, probably via a mechanism similar to that previously described for the E. coli crp gene.