Expression and characterization of Momordica Chanrantia anti-hyperglycaemic peptide in Escherichia coli.

A nucleic acid sequence MC, encoding Momordica Chanrantia anti-hyperglycaemic peptide MC6 (accession: AAX06814) synthesized according to Escherichia coli preferred codons, was cloned and expressed in E. coli. Recombinant protein pQE8-MC (about 3.5 kDa) was purified and analyzed by 20% SDS-PAGE and western blot. It revealed that the expressed pQE8-MC had good solubility in aqueous media. An HPLC assay was used to confirm the expression of pQE8-MC. Subsequent pharmacological activity assay revealed a significant hypoglycemic effect of low dose treatments of pQE8-MC on male kunming mice. Four hours after an intravenous tail injection, the blood sugar levels of mice treated with pQE8-MC saline solution A3 (1 mg/kg BW) decreased greatly (P < 0.01) relative to the levels of a control group. This suggests that pQE8-MC, expressed in bioengineered E. coli, has a similar hypoglycemic function to the natural protein MC6 from M. Chanrantia. These results reveal the possibility of using bio-engineered bacteria as an anti-diabetic agent.

Alga-lysing bioreactor and dominant bacteria strain.

Alga-lysing bacteria have been paid much attention to in recent years. In this study, the alga-lysing strain P05 which was isolated from an immobilizing biosystem was immobilized by coke and elastic filler, forming two biological reactors. The removal efficiencies of algae, NH4(+)-N and organic matter using the two reactors were studied. The results showed that strain P05 was an ideal algal-lysing bacteria strain because it was easy to be immobilized by coke and elastic filler which are of cheap, low biodegradability and the simple immobilization procedure. After 7 d filming, the biological film could be formed and the reactors were used to treat the eutrophic water. These two reactors were of stability and high effect with low cost and easy operation. The optimal hydraulic retention time of each reactor was 4 h. The algae removal rates were 80.38% and 82.1% (in term of Chl-a) of coke reactor and filler reactor, respectively. And that of NH4(+)-N were 52.3% and 52.7%. The removal rates of COD(Mn) were 39.03% and 39.64%. The strain P05 was identified as Bacillus sp. by PCR amplification of the 16S rRNA gene, BLAST analysis, and comparison with sequences in the GenBank nucleotide database.

Isolation and algae-lysing characteristics of the algicidal bacterium B5.

Water blooms have become a worldwide environmental problem. Recently, algicidal bacteria have attracted wide attention as possible agents for inhibiting algal water blooms. In this study, one strain of algicidal bacterium B5 was isolated from activated sludge. On the basis of analysis of its physiological characteristics and 16S rDNA gene sequence, it was identified as Bacillus fusiformis. Its algaelysing characteristics on Microcystis aeruginosa, Chlorella and Scenedesmus were tested. The results showed that: (1) the algicidal bacterium B5 is a Gram-negative bacterium. The 16S rDNA nucleotide sequence homology of strain B5 with 2 strains of B. fusiformis reached 99.86%, so B5 was identified as B. fusiformis; (2) the algal-lysing effects of the algicidal bacterium B5 on M. aeruginosa, Chlorella and Scenedesmus were pronounced. The initial bacterial and algal cell densities strongly influence the removal rates of chlorophyll-a. The greater the initial bacterial cell density, the faster the degradation of chlorophyll-a. The greater the initial algal cell density, the slower the degradation of chlorophyll-a. When the bacterial cell density was 3.6 x 10(7) cells/ml, nearly 90% of chlorophyll-a was removed. When the chlorophyll-a concentration was less than 550 microg/L, about 70% was removed; (3) the strain B5 lysed algae by secreting metabolites and these metabolites could bear heat treatment.

Degradation characteristics of two Bacillus strains on the Microcystis aeruginosa.

The degradation kinetics of strains P05 and P07 and the degradation effects of mixed strain on Microcystis aeruginosa were studied. The results showed that: (1) The degradation processes of strains P05 and P07 on Microcystis aeruginosa accorded with the first-order reaction model when the range of Chl-a concentration was from 0 to 1500 microg/L. (2) The initial bacterium densities had a strong influence on the degradation velocity. The greater the initial bacterium density was, the faster the degradation was. The degradation velocity constants of P05 were 0.1913, 0.2175 and 0.3092 respectively, when bacterium densities were 4.8 x 10(5), 4.8 x 10(6), 2.4 x 10(7) cells/ml. For strain P07, they were 0.1509, 0.1647 and 0.2708. The degradation velocity constant of strain P05 was higher than that of P07 when the bacterium density was under 4.8 x 10(5) cells/ml, but the constant increasing of P07 was quicker than that of P05. (3) The degradation effects of P05 and P07 strains did not antagonize. When the concentration of Chl-a was high, the degradation effects of mixed strain excelled that of any single strains. But with the decrease of the Chl-a concentration, this advantage was not clear. When the concentration was less than 180 microg/L, the degradation effects of mixed were consistent with that of strain P07.