Cloning and heterologous expression of sansanmycin biosynthetic gene cluster in Streptomyces coelicolor / 药学学报

Sansanmycins (SSs), produced by Streptomyces sp. SS, belong to uridyl peptide antibiotics which exhibit a good inhibitory effect on Mycobacterium tuberculosis and Pseudomonas aeruginosa. They share a unique chemical scaffold with a 4',5'-enamide-3'-deoxyuridine attached to DABA (N-methyl-2,3-diaminobutyryl) which was located in the peptide chain through peptide bond. In order to study the function of related genes and to employ synthetic biology to gain new SS derivatives, we obtained a complete SS biosynthetic gene cluster and heterologously expressed it in Streptomyces coelicolor M1146, M1152 and M1154. Fermentation broth of the recombinant strains were detected using HPLC and HPLC-MS/MS, and the result showed that SS-A was successfully produced in the three strains, and its production level in S. coelicolor M1154 was similar to the original wild type strain. In addition, a potential SS analogue named as SS-1154 was discovered from the fermentation broth of S. coelicolor M1154.

Functional analysis of ATP-binding cassette transporter of Streptomyces coelicolor.

ATP- binding cassette (ABC) transporters are characterized as multi drug resistant transporters utilizing ATP hydrolysis. The SCO5113 (GeneID: 1100554) encoding a multidrug resistance ABC transporter in Streptomyces coelicolor was expressed in E.coli BL21 (DE3) plysS, using pET 21 a(+). The expression of his – tag fused SCO5113 at 65 kDa under 0.1 M IPTG in 6 h was detected on SDS-PAGE after purification using affinity chromatography. The E. coli BL21 (DE3) plysS harboring SCO5113 was used to test for the antibiotic resistance, using disc diffussion method. The results showed the resistance of E. coli BL21 (DE3) plysS harboring SCO5113 to five kinds of tested antibiotics such as cephalothin (1mg/ml), kanamycin (1mg/ml), ampicillin (10mg/ml), erythromycin (10mg/ml), and chloramphenicol (1mg/ml). The study reported the function of the SCO5113 gene of Streptomyces coelicolor A3(2).

Studies on the Prodigiosin Production from Streptomyces coelicolor in Liquid Media by Using Heated Lactobacillus rhamnosus.

Prodigiosin (PG) is not only an antibiotic but it has also been characterized immunosuppressive and anticancer activities. Prodigiosin is produced by Streptomyces coelicolor and Seratia marcescens. In this study, prodigiosin (PG) production of S. coelicolor was optimized using 50μL, 100μL of heated cells at 70oC and 100oC of Lactobacillus rhamnosus. With 50μL heat-killed cells at 70oC L. rhamnosus, prodigiosin (PG) production of S. coelicolor enhanced the maximum prodigiosin concentration (9.79  1.68 mg/L) by six-fold at the early stage in the second day incubation in the comparison with prodigiosin (PG) production of S. coelicolor without heated L. rhamnosus (1.43  0.22 mg/L). There were the affects of the components of L. rhamnosus on PG production. This is the first report that L. rhamnosus affecting on PG production in S. coelicolor.

Biodegradation of a blended starch/natural rubber foam biopolymer and rubber gloves by Streptomyces coelicolor CH13

Background: The growing problem of environmental pollution caused by synthetic plastics has led to the search for alternative materials such as biodegradable plastics. Of the biopolymers presently under development, starch/natural rubber is one promising alternative. Several species of bacteria and fungi are capable of degrading natural rubber and many can degrade starch. Results: Streptomyces coelicolor CH13 was isolated from soil according to its ability to produce translucent halos on a mineral salts medium, MSM, supplemented with natural rubber and to degrade starch. Scanning electron microscope studies showed that it colonized the surfaces of strips of a new starch/natural rubber biopolymer and rubber gloves and caused degradation by forming holes, and surface degradation. Starch was completely removed and polyisoprene chains were broken down to produce aldehyde and/or carbonyl groups. After 6 weeks of cultivation with strips of the polymers in MSM, S. coelicolor CH13 reduced the weight of the starch/NR biopolymer by 92 percent and that of the rubber gloves by 14.3 percent. Conclusions: This study indicated that this bacterium causes the biodegradation of the new biopolymer and natural rubber and confirms that this new biopolymer can be degraded in the environment and would be suitable as a ‘green plastic’ derived from natural sources.

Sequence Analysis and Potential Action of Eukaryotic Type Protein Kinase from Streptomyces coelicolor A3(2)

Protein kinase C (PKC) is a family of kinases involved in the transduction of cellular signals that promote lipid hydrolysis. PKC plays a pivotal role in mediating cellular responses to extracellular stimuli involved in proliferation, differentiation and apoptosis. Comparative analysis of the PKC-alpha, beta, epsilon isozymes of 200 recently sequenced microbial genomes was carried out using variety of bioinformatics tools. Diversity and evolution of PKC was determined by sequence alignment. The ser/thr protein kinases of Streptomyces coelicolor A3 (2), is the only bacteria to show sequence alignment score greater than 30% with all the three PKC isotypes in the sequence alignment. S.coelicolor is the subject of our interest because it is notable for the production of pharmaceutically useful compounds including anti-tumor agents, immunosupressants and over two-thirds of all natural antibiotics currently available. The comparative analysis of three human isotypes of PKC and Serine/threonine protein kinase of S.coelicolor was carried out and possible mechanism of action of PKC was derived. Our analysis indicates that Serine/ threonine protein kinase from S. coelicolor can be a good candidate for potent anti-tumor agent. The presence of three representative isotypes of the PKC super family in this organism helps us to understand the mechanism of PKC from evolutionary perspective.

Sequence Analysis and Potential Action of Eukaryotic Type Protein Kinase from Streptomyces coelicolor A3(2)

Protein kinase C (PKC) is a family of kinases involved in the transduction of cellular signals that promote lipid hydrolysis. PKC plays a pivotal role in mediating cellular responses to extracellular stimuli involved in proliferation, differentiation and apoptosis. Comparative analysis of the PKC-alpha, beta, epsilon isozymes of 200 recently sequenced microbial genomes was carried out using variety of bioinformatics tools. Diversity and evolution of PKC was determined by sequence alignment. The ser/thr protein kinases of Streptomyces coelicolor A3 (2), is the only bacteria to show sequence alignment score greater than 30% with all the three PKC isotypes in the sequence alignment. S.coelicolor is the subject of our interest because it is notable for the production of pharmaceutically useful compounds including anti-tumor agents, immunosupressants and over two-thirds of all natural antibiotics currently available. The comparative analysis of three human isotypes of PKC and Serine/threonine protein kinase of S.coelicolor was carried out and possible mechanism of action of PKC was derived. Our analysis indicates that Serine/ threonine protein kinase from S. coelicolor can be a good candidate for potent anti-tumor agent. The presence of three representative isotypes of the PKC super family in this organism helps us to understand the mechanism of PKC from evolutionary perspective.