Distribution of Bacterial Communities in Petroleum-Contaminated Soils from the Dagang Oilfield, China

Diversity in bacterial communities was investigated along a petroleum hydrocarbon content gradient(0-0.4043 g/g)in surface(5-10 cm)and subsurface(35-40 cm)petroleum-contaminated soil samples from the Dagang Oilfield,China.Using 16S rRNA Illumina high-throughput sequencing technology and several statistical methods,the bacterial diversity of the soil was studied.Subsequently,the environmental parameters were measured to analyze its relationship with the community variation.Nonmetric multidimensional scaling and analysis of similarities indicated a significant difference in the structure of the bacterial community between the nonpetroleum-contaminated surface and subsurface soils,but no differences were observed in different depths of petroleum-contaminated soil.Meanwhile,many significant correlations were obtained between diversity in soil bacterial community and physicochemical properties.Total petroleum hydrocarbon,total organic carbon,and total nitrogen were the three important factors that had the greatest impacts on the bacterial community distribution in the long-term petroleum-contaminated soils.Our research has provided references for the bacterial community distribution along a petroleum gradient in both surface and subsurface petroleum-contaminated soils of oilfield areas.

Research of 1,3-Dihydroxyacetone Production by Overexpressing Glycerol Transporter and Glycerol Dehydrogenase

1,3-Dihydroxyacetone (DHA), a natural ketose, is widely used in the chemical, cosmetic, and pharmaceutical industries. The current method for DHA production is Gluconobacter oxydans ( G. oxydans ) fermentation, but the high concentration of glycerol in the fermentation broth inhibits cells growth. To overcome this obstacle, in this study, we overexpressed the glycerol transporter (GlpFp) by the use of promoters P tufB , P gmr , P glp1 , and P glp2 in G. oxydans 621H. The results show that the glycerol tolerances of strains overexpressing G lpF were all much better than that of the control strain. The glycerol dehydrogenase gene (G dh) was overexpressed by the promoters P tufB and P gdh , which increased the DHA titer by 12.7% compared with that of the control group. When G lpF and Gdh genes were co-overexpressed in G. oxydans 621H, the OD600 value of the engineered strains all increased, but the DHA titers decreased in di erent degrees, as compared with strains that overexpressed only G dh . This study provides a reference for future research on DHA production.

Selection of Reference Genes in Saccharopolyspora Spinosa for Real-Time PCR

Reverse transcription quantitative PCR （RT-qPCR） combined with the published genome information of Saccharopolyspora spinosa can allow sophisticated studies about S. spinosa, including Studying the regulation of spinosyn biosynthesis, finding new target genes for engineering, and discovering and exploiting other macrolide secondary metabolites. Studies have demonstrated that appropriate internal control is needed to normalize target genes at transcription levels. However, many studies have shown that no single reference gene is universal for all strains under all experimental conditions. Thus, eight candidate reference genes of three different S. spinosa strains in two different cultures were studied to find suitable reference gene（sl. The number of amplification cycles of these candidate genes was calculated by BestKeeper, NormFinder and geNorm. The results indicated that the most suitable reference genes for normalization during the fermentation of S. spinosa were 16S rRNA and rbL13.

Engineering Corynebacterium glutamicum for Geraniol Production

Geraniol is a monoterpenoid alcohol with various applications in food,cosmetics,and healthcare.Corynebacterium glutamicum is a potential platform for terpenoids production because it harbors the methylerythritol phosphate pathway.To engineer C.glutamicum to produce geraniol,two different truncated geraniol synthases (GESs) were respectively expressed,and strain LX02 expressing the truncated GESs from Valeriana officinalis (t Vo GES) produced 0.3 mg/L of geraniol.Then,three geranyl diphosphate synthases (GPPSs) were combinatorially co-expressed with t Vo GES to improve geraniol production.The amounts of produced geraniol were all higher than that produced by strain LX02.Strain LX03 co-expressing ERG20 F96W–N127W (ERG20 WW) and t Vo GES produced the highest amount,5.4 mg/L.Subsequently,the co-overexpression of1-deoxy-D-xylulose-5-phosphate synthase (dxs) and isopentenyl diphosphate isomerase (idi) further increased the production to 12.2 mg/L in strain LX03.Lastly,the production of geraniol was increased to 15.2 mg/L via fermentation optimization.To our knowledge,this is the first report on the engineering of C.glutamicum to produce geraniol and thus can serve as a reference for other monoterpenoid production studies.