Microbial production of mevalonate.

Mevalonate, an important intermediate product of the mevalonate pathway, has a broad spectrum of applications. With the rapid growth of metabolic engineering and synthetic biology, mevalonate biosynthesis by microorganisms is feasible and holds great promise in the future. In this review, we summarize the applications of mevalonate and its derivatives and describe the biosynthesis pathways of mevalonate. The current status of mevalonate biosynthesis is also detailed with an emphasis on metabolic engineering strategies to enhance mevalonate production in typical industrial organisms, including Escherichia coli, Saccharomyces cerevisiae, and Pseudomonas putida, suggesting new insights for the efficient production of biosynthesized mevalonate.

Sulfur metabolism in Rhodococcus species and their application in desulfurization of fossil fuels.

Organosulfur compounds in fossil fuels have been a major concern in the process of achieving zero-sulfur fuel production. Biodesulfurization (BDS) is an environmentally friendly strategy for the removal of refractory organosulfur compounds from fossil fuels. Even though researchers are committed to engineering the desulfurization-specific pathway for improving BDS efficiency, the industrial application of BDS is still difficult. Recently, the sulfur metabolism of Rhodococcus has begun to attract attention due to its influences on the BDS process. In this review, we introduce the sulfur metabolism in Rhodococcus, including sulfur absorption, reduction, and assimilation; and summarize desulfurization in Rhodococcus, including the desulfurization mechanism, the regulation mechanism of the 4S pathway, and the strategies of optimizing the 4S pathway to improve BDS efficiency. In particular, the influence of sulfur metabolism on BDS efficiency is discussed. In addition, we consider the latest genetic engineering strategies in Rhodococcus. An improved understanding of the relationship between sulfur metabolism and desulfurization will enable the industrial application of BDS.

Synthetic small regulatory RNAs in microbial metabolic engineering.

Small regulatory RNAs (sRNAs) finely control gene expression in prokaryotes and synthetic sRNA has become a useful high-throughput approach to tackle current challenges in metabolic engineering because of its many advantages compared to conventional gene knockouts. In this review, we first focus on the modular structures of sRNAs and rational design strategies of synthetic sRNAs on the basis of their modular structures. The wide applications of synthetic sRNAs in bacterial metabolic engineering, with or without the aid of heterogeneously expressed Hfq protein, were also covered. In addition, we give attention to the improvements in implementing synthetic sRNAs, which make the synthetic sRNA strategy universally applicable in metabolic engineering and synthetic biology. KEY POINTS: • Synthetic sRNAs can be rationally designed based on modular structures of natural sRNAs. • Synthetic sRNAs were widely used for metabolic engineering in various microorganisms. • Several technological improvements made the synthetic sRNA strategy more applicable.

Molecular epidemiological survey of bacterial and parasitic pathogens in hard ticks from eastern China.

Ticks are able to transmit various pathogens-viruses, bacteria, and parasites-to their host during feeding. Several molecular epidemiological surveys have been performed to evaluate the risk of tick-borne pathogens in China, but little is known about pathogens circulating in ticks from eastern China. Therefore, this study aimed to investigate the presence of bacteria and parasites in ticks collected from Xuzhou, a 11258km2 region in eastern China. In the present study, ticks were collected from domestic goats and grasses in urban districts of Xuzhou region from June 2015 to July 2016. After tick species identification, the presence of tick-borne bacterial and parasitic pathogens, including Anaplasma phagocytophilum, Borrelia burgdorferi, Rickettsia sp., Bartonella sp., Babesia sp., and Theileria sp., was established via conventional or nested polymerase chain reaction assays (PCR) and sequence analysis. Finally, a total of 500 questing adult ticks, identified as Haemaphysalis longicornis, were investigated. Among them, 28/500 tick samples (5.6%) were infected with A. phagocytophilum, and 23/500 (4.6%) with Theileria luwenshuni, whereas co-infection with these pathogens was detected in only 1/51 (2%) of all infected ticks. In conclusion, H. longicornis is the dominant tick species in the Xuzhou region and plays an important role in zoonotic pathogen transmission. Both local residents and animals are at a significant risk of exposure to anaplasmosis and theileriosis, due to the high rates of A. phagocytophilum and T. luwenshuni tick infection.

Role of the spleen in Bartonella spp. infection.

Bartonella spp. are intra-erythrocytic pathogens of mammals. In this study, we investigated the role of the spleen, and other tissue and organs in Bartonella infection. Using an in vivo model of mice infection by Bartonella birtlesii, we detected accumulation of bacteria in the spleen, with transient infection of the liver, but failed to detect any bacteria in brain or lymph nodes. We then compared bacteraemia in normal Balb/C mice and in splenectomized mice. Bacteraemia in splenectomized mice was 10-fold higher than in normal mice and lasted 2 weeks longer. In conclusion, the spleen seems to retain and filter infected erythrocytes rather than to be a sanctuary for chronic Bartonella infection.

The Trw type IV secretion system of Bartonella mediates host-specific adhesion to erythrocytes.

Bacterial pathogens typically infect only a limited range of hosts; however, the genetic mechanisms governing host-specificity are poorly understood. The alpha-proteobacterial genus Bartonella comprises 21 species that cause host-specific intraerythrocytic bacteremia as hallmark of infection in their respective mammalian reservoirs, including the human-specific pathogens Bartonella quintana and Bartonella bacilliformis that cause trench fever and Oroya fever, respectively. Here, we have identified bacterial factors that mediate host-specific erythrocyte colonization in the mammalian reservoirs. Using mouse-specific Bartonella birtlesii, human-specific Bartonella quintana, cat-specific Bartonella henselae and rat-specific Bartonella tribocorum, we established in vitro adhesion and invasion assays with isolated erythrocytes that fully reproduce the host-specificity of erythrocyte infection as observed in vivo. By signature-tagged mutagenesis of B. birtlesii and mutant selection in a mouse infection model we identified mutants impaired in establishing intraerythrocytic bacteremia. Among 45 abacteremic mutants, five failed to adhere to and invade mouse erythrocytes in vitro. The corresponding genes encode components of the type IV secretion system (T4SS) Trw, demonstrating that this virulence factor laterally acquired by the Bartonella lineage is directly involved in adherence to erythrocytes. Strikingly, ectopic expression of Trw of rat-specific B. tribocorum in cat-specific B. henselae or human-specific B. quintana expanded their host range for erythrocyte infection to rat, demonstrating that Trw mediates host-specific erythrocyte infection. A molecular evolutionary analysis of the trw locus further indicated that the variable, surface-located TrwL and TrwJ might represent the T4SS components that determine host-specificity of erythrocyte parasitism. In conclusion, we show that the laterally acquired Trw T4SS diversified in the Bartonella lineage to facilitate host-restricted adhesion to erythrocytes in a wide range of mammals.

[Limitations of the application of RNA interference in the study of animal parasitic nematodes].

RNA interference is a powerful tool for investigating gene function which has been used extensively in model organisms such as Caenorhabditis elegans and has been adapted as a high-throughput screening method to identify genes. However, recent research indicates that the application of RNAi to animal parasitic nematodes has some problems. For example, the variability of RNAi to different parasitic nematodes or the same nematode in different stages is great. Reasons to explain why RNAi does not work well in animal parasitic nematodes include: (1) the efficacy of RNAi delivery methods in different nematodes is extremely variable; (2) RNAi mechanism has been applied to related nematodes and most of them are gene functional defect; (3) different lifestyles of nematodes may influence on the efficacy of RNAi. If we can find out the methods which can solve these questions, there are still application prospects for using RNAi in parasitic nematodes with improved RNAi effect.