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The low enzymatic capability of terpene synthases and the limited availability of precursors often hinder the productivity of terpenes in microbial hosts. Herein, a systematic approach combining protein engineering and pathway compartmentation was exploited in

The traditional channel sounding protocol requires that the whole sounding process must be completed within a coherent time. When the communication delay is close to or greater than the coherent time, the protocol will not work properly. In order to solve this problem, a novel Non-Time-Limited channel sounding protocol is proposed. The proposed channel sounding protocol does not need to guarantee the channel reciprocity, has no limit on the communication delay and has strong multi-relay scalability. At the same time, a relay-assisted security extension mechanism is proposed to prevent the collaborative eavesdropping of the neighboring attackers. Simulation experiments show that the Non-Time-Limited channel sounding protocol can get rid of the influence of channel change speed on key generation, effectively improve key consistency in high-speed mobile scenarios, and still maintain good key agreement rate performance in underwater acoustic communication.

α-Pinene, an important biologically active natural monoterpene, has been widely used in fragrances, medicines, and fine chemicals, especially, in high-density renewable fuels such as jet fuel. The development of an α-pinene production platform in a highly modifiable microbe from renewable substitute feedstocks could lead to a green, economical avenue, and sustainable biotechnological process for the biosynthesis of α-pinene. Here, we report engineering of an orthogonal biosynthetic pathway for efficient production of α-pinene in oleaginous yeast Yarrowia lipolytica that resulted in an α-pinene titer of 19.6 mg/L when using glucose as the sole carbon source, a significant 218-fold improvement than the initial titer. In addition, the potential of using waste cooking oil and lignocellulosic hydrolysate as carbon sources for α-pinene production from the engineered Y. lipolytica strains was analyzed. The results indicated that α-pinene titers of 33.8 and 36.1 mg/L were successfully obtained in waste cooking oil and lignocellulosic hydrolysate medium, thereby representing the highest titer reported to date in yeast. To our knowledge, this is also the first report related to microbial production of α-pinene from waste cooking oil and lignocellulosic hydrolysate.

The oleaginous yeast Yarrowia lipolytica is an attractive cell factory platform strain and can be used for sustainable production of high-value oleochemical products. Wax esters (WEs) have a good lubricating property and are usually used as a base for the production of advanced lubricants and emollient oils. In this study, we reported the metabolic engineering of Y. lipolytica to heterologously biosynthesize high-content very-long-chain fatty acids (VLCFAs) and fatty alcohols and efficiently esterify them to obtain very-long-chain WEs. Co-expression of fatty acid elongases from different sources in Y. lipolytica could yield VLCFAs with carbon chain lengths up to 24. Combining with optimization of the central metabolic modules could further enhance the biosynthesis of VLCFAs. Furthermore, through the screening of heterologous fatty acyl reductases (FARs), we enabled high-level production of fatty alcohols. Genomic integration and heterologous expression of wax synthase (WS) and FAR in a VLCFA-producing Y. lipolytica strain yielded 95-650 mg/L WEs with carbon chain lengths from 32 to 44. Scaled-up fermentation in 5 L laboratory bioreactors significantly increased the production of WEs to 2.0 g/L, the highest content so far in yeasts. This study contributes to the further efficient biosynthesis of VLCFAs and their derivatives.

Farnesene is a typical sesquiterpene with applications as fragrance, flavor and precursor for the synthesis of vitamin E/K1. In this study, a series of strategies were employed to facilitate α-farnesene accumulation in Yarrowia lipolytica. Among them, the promoter optimization of OptFSLERG20, Sc-tHMG1 and IDI resulted in more than 62 % increase in α-farnesene production. Together with the overexpression of Yl-HMGR and ERG19, α-farnesene content was significantly improved by more than 3.5 times. The best metabolic engineered strain obtained was therefore used for a uniform design in shake flasks to determine the optimal medium compositions. Furthermore, a maximum α-farnesene production of approximately 2.57 g/L (34 mg/g DCW) was obtained in fed-batch fermentation where glycerol was supplemented as the feeding carbon source when initial glucose was depleted. This study has laid a good foundation for the development of Y. lipolytica as a promising chassis microbial cell for heterologous biosynthesis of α-farnesene and other sesquiterpenes.

Ansamitocin P-3 (AP-3) shows strong anticancer effects and has used as a payload for antibody-drug conjugates. Our previous study have shown that although genetically engineered Actinosynnema pretiosum strains with enhanced UDP-glucose (UDPG) biosynthesis displayed improved AP-3 production compared to the wild-type strain, the increase in yield was far from meeting the industrial demand. In this study, comparative metabolomics analysis complemented with quantitative real-time PCR analysis was performed for the wild-type strain and two mutants (OpgmOugp, ΔzwfΔgnd) to identify possible metabolic bottlenecks and non-intuitive targets for further enhancement of AP-3 production. We observed that enhancing intracellular UDPG availability facilitated the accumulation of intracellular N-demethyl-AP-3 and AP-3, where the transporting of them outside the cell still needs to be developed. We also found that the UDPG biosynthesis was closely associated with the availability of fructose in the medium and a suitable fructose feeding strategy could promote the further improvement of AP-3 titer. In addition, pathway abundance analysis revealed that undesired fatty acid accumulation and down-regulation of amino acid metabolism may be unfavorable for ansamitocin biosynthesis in later stage of production. These results indicate that genetic modification of the UDPG biosynthetic pathways may have pleiotropic effects on AP-3 production. Efforts must be made to eliminate these newly identified metabolic bottlenecks to boost AP-3 production in A. pretiosum.

The CRA-Interim trial production of the global atmospheric reanalysis for 10 years from 2007 to 2016 was carried out by the China Meteorological Administration in 2017. The structural characteristics of the horizontal shear line over the Tibetan Plateau (TPHSL) based on the CRA-Interim datasets are examined by objectively identifying the shear line, and are compared with the analysis results of the European Centre for Medium-Range Weather Forecasts reanalysis data (ERA-Interim). The case occurred at 18UTC on July 5, 2016. The results show that both of the ERA-Interim and CRA-Interim datasets can well reveal the circulation background and the dynamic and thermal structure characteristics of TPHSL, and they have shown some similar features. The middle and high latitudes at 500 hPa are characterized by the circulation situation of"two troughs and two ridges", and at 200 hPa, the TPHSL is located in the northeast quadrant of the South Asian High Pressure (SAHP). The TPHSL locates in the positive vorticity zone and passes through the positive vorticity center corresponding to the ascending motion. Near the TPHSL, the contours of pseudo-equivalent potential temperature (θse) tend to be intensive, with a high-value center on the south side of the TPHSL. The TPHSL can extend to 460 hPa and vertically inclines northward. There is a positive vorticity zone near the TPHSL which is also characterized by the northward inclination with the height, the ascending motion near the TPHSL can extend to 300 hPa, and the atmospheric layer above the TPHSL is stable. However, the intensities of the TPHSL's structure characteristics analyzed with the two datasets are different, revealing the relatively strong intensity of geopotential height field, vertical velocity field, vorticity field and divergence field from the CRA-Interim datasets. In addition, the vertical profiles of the dynamic and water vapor thermal physical quantities of the two datasets are also consistent in the east and west part of the TPHSL. In summary, the reliable and usable CRA-Interim datasets show excellent properties in the analysis on the structural characteristics of a horizontal shear line over the Tibetan Plateau.