Microbial cell factories (bacteria and fungi) are the leading producers of beneficial natural products such as lycopene, carotene, herbal medicine, and biodiesel etc. These microorganisms are considered efficient due to their effective bioprocessing strategy (monoculture- and consortial-based approach) under distinct processing conditions. Meanwhile, the advancement in genetic and process optimization techniques leads to enhanced biosynthesis of natural products that are known functional ingredients with numerous applications in the food, cosmetic and medical industries. Natural consortia and monoculture thrive in nature in a small proportion, such as wastewater, food products, and soils. In similitude to natural consortia, it is possible to engineer artificial microbial consortia and program their behaviours via synthetic biology tools. Therefore, this review summarizes the optimization of genetic and physicochemical parameters of the microbial system for improved production of natural products. Also, this review presents a brief history of natural consortium and describes the functional properties of monocultures. This review focuses on synthetic biology tools that enable new approaches to design synthetic consortia; and highlights the syntropic interactions that determine the performance and stability of synthetic consortia. In particular, the effect of processing conditions and advanced genetic techniques to improve the productibility of both monoculture and consortial based systems have been greatly emphasized. In this context, possible strategies are also discussed to give an insight into microbial engineering for improved production of natural products in the future. In summary, it is concluded that the coupling of genomic modifications with optimum physicochemical factors would be promising for producing a robust microbial cell factory that shall contribute to the increased production of natural products.

The stability and high productivity of heterogeneous terpenoid production in Escherichia coli expression system is one of the most key issues for its large scale industrialization. In the current study on taking lycopene biosynthesis as an example, an integrated Escherichia coli system has been generated successfully, which resulted into stable and high lycopene production. In this process, two modules of mevalonate (MVA) pathway and one module of lycopene expression pathway were completely integrated in the chromosome. Firstly, the copy number and integrated position of three modules of heterologous pathways were rationally optimized. Later, a strain DH416 equipped with heterogeneous expression pathways through chromosomal integration was efficiently derived from parental strain DH411. The evolving DH416 strain efficiently produced the lycopene level of 1.22 g/L (49.9 mg/g DCW) in a 5 L fermenter with mean productivity of 61.0 mg/L/h. Additionally, the integrated strain showed more genetic stability than the plasmid systems after successive 21st passage.

BACKGROUND:RNA interference (RNAi)-based pest management requires efficient delivery and large-batch production of double-stranded (ds)RNA. We previously developed a nanocarrier-mediated dsRNA delivery system that could penetrate an insect's body and efficiently silence gene expression. However, there is a great need to improve the plasmid-Escherichia coli system for the mass production of dsRNA. Here, for efficient dsRNA production, we removed the rnc gene encoding endoribonuclease RNase III in E. coli BL21(DE3) and matched with the RNAi expression vector containing a single T7 promoter. RESULTS:The novel pET28-BL21(DE3) RNase III-system was successfully constructed to express vestigial (vg)-dsRNA against Harmonia axyridis. dsRNA was extracted and purified from cell cultures in four E. coil systems, and the yields of dsRNA in pET28-BL21(DE3) RNase III-, pET28-HT115(DE3), L4440-BL21(DE3) RNase III- and L4440-HT115(DE3) were 4.23, 2.75, 0.88 and 1.30 μg mL-1 respectively. The dsRNA expression efficiency of our novel E. coil system was three times that of L4440-HT115(DE3), a widely used dsRNA production system. The RNAi efficiency of dsRNA produced by our system and by biochemical synthesis was comparable when injected into Harmonia axyridis. CONCLUSION:Our system expressed dsRNA more efficiently than the widely used L4440-HT115(DE3) system, and the produced dsRNA showed a high gene-silencing effect. Notably, our pET28-BL21(DE3) RNase III-system provides a novel method for the mass production of dsRNA at low cost and high efficiency, which may promote gene function analysis and RNAi-based pest management. © 2020 Society of Chemical Industry.

Camel milk is worldwide known for its profound nutritional value. However, the poor curd formation of camel milk renders low consumer acceptability. In current study, xanthan gum was added in the production process to attain an adequate firmness factor. Further for enhancing consumer acceptability, dates paste was also added. Storage study was performed four times after an elapse of every 7 days to confirm the product quality for corresponding fractions (0.25, 0.5 and 0.75%) of biosynthesized xanthan (BX) and 1% gelatin as control. Moreover, the rheological measurements were performed and the experimental data were assessed by Herschel-Bulkley model. With the addition of BX, the camel milk date yoghurt (CMDY) demonstrated the shear-thinning behavior with pseudoplastic fluid characteristics. The rheological properties of CMDY were significantly improved in correspondence to higher concentration of BX. Relatively, 0.75% of BX addition resulted in the best texture of yoghurt and obtained an average firmness of 445 +/- 34.11 g. Furthermore, confocal CSLM and SEM micrographs demonstrated the detail microstructure of all yoghurt samples. Specifically, addition of 0.75% BX in yoghurt sample demonstrated a strengthened (dense) network of casein micelles. Lastly, the addition of 0.75% BX along with 10% dates paste significantly enhanced the sensory characteristics of CMDY.

[This corrects the article DOI: 10.1371/journal.pone.0186891.].

The multiple plasmid system, mostly relied, for heterogeneous gene expression, results in genetic instability and low mean productivity. To address this, an integration method was employed for investigating expression of heterogenous pathway in E. coli cells; where mevalonate upper pathway was found efficiently expressed. Subsequently, to improve lycopene production, chromosomal multiple position integration strategy was used to strengthen mevalonate upper pathway. Meanwhile, the plasmid system was employed for mevalonate lower pathway and lycopene pathway expression to finally generate the mutant D711 strain. Comparatively, highest level of 68.5 mg/L lycopene was produced by D711 outperforming its maximum average productivity of 2.85 mg/L/h with over 2-folds enhancement. In addition, lycopene level was almost 224 mg/L after optimization of induction time, which was 3.3-fold higher than standard control condition. Finally, expression Performance Parameter was developed for scoring mutants and evaluating these two strategies, indicating chromosomal multiple position integration strategy as more efficient approach.

Objective: To apply a complementary resistance strategy to increase the in vivo direct cloning efficien?cy mediated by Red recombination. Methods: The cat cassette was divided into two parts, Cma and Cmb(has a homologous sequence with Cma), each of them has no resistance separately unless the two parts are fused togeth?er. The Cmb fragment was first integrated into a side of the target cloning region, then a linear Cma-containing plasmid vector was electroporated into the cells to directly capture the region. Results: Based on this strategy, we successfully cloned an about 48 kb DNA fragments from E.coli DH1 chromosome. Conclusion: We can use this method in functional studies of large DNA.