Recent advances in the molecular genetics of resin biosynthesis and genetic engineering strategies to improve defenses in conifers

Since the first terpenoid synthase cDNA was obtained by the reverse genetic approach from grand fir, great progress in the molecular genetics of terpenoid formation has been made with angiosperms and genes encoding a monoterpene synthase, a sesquiterpene synthase, and a diterpene synthase. Tree killing bark beetles and their vectored fungal pathogens are the most destructive agents of conifer forests worldwide. Conifers defend against attack by the constitutive and inducible production of oleoresin that accumulates at the wound site to kill invaders and both flush and seal the injury. Although toxic to the bark beetle and fungal pathogen, oleoresin also plays a central role in the chemical ecology of these boring insects. Recent advances in the molecular genetics of terpenoid biosynthesis provide evidence for the evolutionary origins of oleoresin and permit consideration of genetic engineering strategies to improve conifer defenses as a component of modern forest biotechnology. This review described enzymes of resin biosynthesis, structural feathers of genes genomic intron and exon organization, pathway organization and evolution, resin production and accumulation, interactions between conifer and bark beetle, and engineering strategies to improve conifer defenses.

Metabolic engineering strategies for microbial utilization of methanol

The increasing shortage of fossil resources and environmental pollution has renewed interest in the synthesis of value-added biochemicals from methanol.However,most of native or synthetic methylotrophs are unable to assimilate methanol at a sufficient rate to produce biochemicals.Thus,the performance of methylotrophs still needs to be optimized to meet the demands of industrial applications.In this review,we provide an in-depth discussion on the properties of natural and synthetic methylotrophs,and summarize the natural and synthetic methanol assimilation pathways.Further,we discuss metabolic engineering strategies for enabling microbial utilization of methanol for the bioproduction of value-added chemicals.Finally,we highlight the potential of microbial engineering for methanol assimilation and offer guidance for achieving a low-carbon footprint for the biosynthesis of chemicals.

Metabolic engineering strategies for microbial utilization of C1 feedstocks

The use of abundant and cheap one carbon(C1)feedstocks to produce value-added chemicals is an important approach for achieving carbon neutrality and tackling environmental problems.The conversion of C1 feedstocks to high-value chemicals is dependent on efficient C1 assimilation pathways and microbial chassis adapted for efficient incorporation.Here,we opted to summarize the natural and synthetic C1 assimilation pathways and their key factors for metabolizing C1 feedstock.Accordingly,we discussed the metabolic engineering strategies for enabling the microbial utilization of C1 feedstocks for the bioproduction of value-added chemicals.In addition,we highlighted future perspectives of C1-based biomanufacturing for achieving a low-carbon footprint for the biosynthesis of chemicals.

Recent Strategies for the Development of Biosourced-Monomers, Oligomers and Polymers-Based Materials: A Review with an Innovation and a Bigger Data Focus

After setting the ground of the quantum innovation potential of biosourced entities and outlining the inventive spectrum of adjacent technologies that can derive from those, the current review highlights, with the support of Bigger Data approaches, and a fairly large number of articles, more than 250 and 10,000 patents, the following. It covers an overview of biosourced chemicals and materials, mainly biomonomers, biooligomers and biopolymers;these are produced today in a way that allows reducing the fossil resources depletion and dependency, and obtaining environmentally-friendlier goods in a leaner energy consuming society. A process with a realistic productivity is underlined thanks to the implementation of recent and specifically effective processes where engineered microorganisms are capable to convert natural non-fossil goods, at industrial scale, into fuels and useful high-value chemicals in good yield. Those processes, further detailed, integrate: metabolic engineering involving 1) system biology, 2) synthetic biology and 3) evolutionary engineering. They enable acceptable production yield and productivity, meet the targeted chemical profiles, minimize the consumption of inputs, reduce the production of by-products and further diminish the overall operation costs. As generally admitted the properties of most natural occurring biopolymers (e.g., starch, poly (lactic acid), PHAs.) are often inferior to those of the polymers derived from petroleum;blends and composites, exhibiting improved properties, are now successfully produced. Specific attention is paid to these aspects. Then further evidence is provided to support the important potential and role of products deriving from the biomass in general. The need to enter into the era of Bigger Data, to grow and increase the awareness and multidimensional role and opportunity of biosourcing serves as a conclusion and future prospects. Although providing a large reference database, this review is largely initiatory, therefore not mimicking previous classic reviews but putting them in a multiplying synergistic prospective.

Metabolic Engineering of Tropane Alkaloid Biosynthesis in Plants

Abstract: Over the past decade, the evolving commercial importance of so-called plant secondary metabolites has resulted in a great interest in secondary metabolism and, particularly, in the possibilities to enhance the yield of fine metabolites by means of genetic engineering. Plant alkaloids, which constitute one of the largest groups of natural products, provide many pharmacologically active compounds. Several genes in the tropane alkaloids biosynthesis pathways have been cloned, making the metabolic engineering of these alkaloids possible. The content of the target chemical scopolamine could be significantly increased by various approaches, such as introducing genes encoding the key biosynthetic enzymes or genes encoding regulatory proteins to overcome the specific rate-limiting steps. In addition, antisense genes have been used to block competitive pathways. These investigations have opened up new, promising perspectives for increased production in plants or plant cell culture. Recent achievements have been made in the metabolic engineering of plant tropane alkaloids and some new powerful strategies are reviewed in the present paper.

Cell-free protein synthesis enabled rapid prototyping for metabolic engineering and synthetic biology

Advances in metabolic engineering and synthetic biology have facilitated the manufacturing of many valuable-added compounds and commodity chemicals using microbial cell factories in the past decade.However,due to complexity of cellular metabolism,the optimization of metabolic pathways for maximal production represents a grand challenge and an unavoidable barrier for metabolic engineering.Recently,cell-free protein synthesis system(CFPS)has been emerging as an enabling alternative to address challenges in biomanufacturing.This review summarizes the recent progresses of CFPS in rapid prototyping of biosynthetic pathways and genetic circuits(biosensors)to speed up design-build-test(DBT)cycles of metabolic engineering and synthetic biology.

Genetic tools for metabolic engineering of Pichia pastoris

The methylotrophic yeast Pichia pastoris(also known as Komagataella phaffii)is widely used as a yeast cell factory for producing heterologous proteins.Recently,it has gained attention for its potential in producing chemicals from inexpensive feedstocks,which requires efficient genetic engineering platforms.This review provides an overview of the current advances in developing genetic tools for metabolic engineering of P.pastoris.The topics cover promoters,terminators,plasmids,genome integration sites,and genetic editing systems,with a special focus on the development of CRISPR/Cas systems and their comparison to other genome editing tools.Additionally,this review highlights the prospects of multiplex genome integration,fine-tuning gene expression,and single-base editing systems.Overall,the aim of this review is to provide valuable insights into current genetic engineering and discuss potential directions for future efforts in developing efficient genetic tools in P.pastoris.

生物合成法生产麦角硫因的研究进展

麦角硫因(ergothioneine,ERG)作为一种稀有的天然含硫组氨酸衍生物,已被证明具有强大的抗氧化性和诸多生物学功能。因此,ERG受到越来越多研究人员和产品开发人员的关注。目前,ERG已被广泛应用于食品、化妆品和医疗等行业。研究表明只有少数细菌和真菌可体内合成ERG,植物、动物和人类自身均不能直接合成ERG,只能从其他来源获取。ERG可通过生物提取法、化学合成法以及生物合成法获得,但由于传统生产方式(生物提取法和化学合成法)存在产量低、生产效率差和生产成本较高等问题,限制了该产品的规模化生产和应用。因此,亟需开发一种高效、经济且安全、可靠的ERG生产方式以满足市场的需求。近年来合成生物学快速发展,利用基因工程、蛋白质工程和代谢工程等技术提高生物合成法生产ERG的能力已逐渐成为研究热点。本文将论述ERG的生物学活性和功能,介绍ERG生物合成途径和ERG在食品、化妆品和医疗等行业的应用前景,比较ERG主要的生产方式,总结并梳理近年来采取各种工程策略通过生物合成法生产ERG的研究进展;并就如何利用基因工程、蛋白质工程和代谢工程提高ERG产量提出几点工程策略,以期为生物合成法高产ERG提供理论参考和研究思路。

微藻破囊壶菌产功能性脂肪酸DHA研究进展

二十二碳六烯酸(DHA)作为人体必需的多不饱和脂肪酸,在维护心血管健康、抗癌、支持视觉和脑功能等方面至关重要。传统的深海鱼油提取DHA方法存在鱼腥味重、工艺繁琐等问题,迫使研究者寻求更为高效、环保的替代方案。破囊壶菌(Thraustochytrids)凭借其生长迅速、低重金属污染以及高DHA含量的特性,成为工业化生产DHA的潜力微生物之一。当前在破囊壶菌发酵生产DHA的过程中,依然需要解决一系列关键问题,包括提高发酵产量、降低成本等。本文旨在全面阐述破囊壶菌发酵生产DHA的研究现状,包括菌株筛选与改良、DHA生物合成途径、遗传转化及代谢工程、发酵控制策略等方面。首先,总结归纳了对野生型菌株的自然筛选和诱变改良等方法,不断提高破囊壶菌中DHA产油量。其次,详细介绍了破囊壶菌DHA合成途径的研究进展,着重分析了生物合成途径中关键辅助因子在DHA生产中的作用。此外,概述了外源DNA传递到破囊壶菌细胞的遗传转化技术的应用现状,为提高其遗传转化效率和稳定性提供重要参考。在DHA代谢调控方面,探讨了氮限制对DHA合成的促进作用以及温度和氧气供应对生产效率的影响。最后,对利用破囊壶菌生产DHA存在的主要瓶颈问题和未来发展趋势进行了总结,以推动其在医药、保健品和食品等领域的广泛应用,实现工业规模下的高效生产。

基于非天然氨基酸的蛋白质合成研究进展

蛋白质是生命体的重要组成部分,几乎参与了所有的生命活动。通常情况下生物体中蛋白质由20种天然氨基酸组成,但随着现代科技的飞速发展,人们对医疗、食品、材料等方面的需求日渐增大,这些蛋白质已经无法满足多样化的应用需求。随着非天然氨基酸及其合成的蛋白质的优异性能,使得利用非天然氨基酸改造蛋白质的技术逐渐成为研究热点。非天然氨基酸有着不同于天然氨基酸的侧链基团,可以给蛋白质带来新的性质与功能。该文基于对非天然氨基酸及非天然蛋白质的研究,首先介绍了以化学及生物的手段合成非天然氨基酸的方法,同时对化学及生物法的优劣势及未来的发展前景进行了总结;然后介绍了将非天然氨基酸掺入蛋白质的几种热门方法;接着对非天然氨基酸及其合成蛋白质的应用在酶的改造、科研以及医疗方面进行了列举和描述;最后对这项技术未来的发展进行了展望与总结。

重组人糖蛋白激素β5/α2融合蛋白在CHO-S细胞中的表达纯化及功能活性分析

目的在悬浮中国仓鼠卵巢细胞(Chinese hamster ovary cells,CHO-S)中分泌表达、纯化重组hCGH-CTP融合蛋白,验证其对3T3-L1成熟脂肪细胞脂质积累的影响。方法构建CTP连接肽融合人糖蛋白激素β5/α2重组蛋白表达载体pcDNA3.1-rhCGH-CTP,将其瞬时转染CHO-S悬浮细胞中,大量表达纯化并验证rhCGH-CTP蛋白生物学活性;通过干预3T3-L1成熟脂肪细胞24 h,观察细胞内甘油三酯(TG)水平的变化。结果Western blot结果显示,rhCGH-CTP蛋白在CHO-S细胞中成功表达,表达量可达715.4 mg·L^(-1);用AKTA pure蛋白纯化系统纯化蛋白,SDS-PAGE方法鉴定纯化出的蛋白纯度较高可达90%。此外,在高表达TSHR基因的成熟脂肪细胞3T3-L1中,利用ELISA试剂盒测定不同浓度rhCGH-CTP蛋白干预后胞内cAMP含量明显升高,说明rhCGH-CTP蛋白具有生物活性;油红O染色结果发现,与对照组相比,不同浓度rhCGH-CTP蛋白干预组的成熟脂肪细胞中TG含量明显降低(P<0.05)。结论成功表达并纯化了rhCGH-CTP融合蛋白,其具有良好的生物学活性并能有效降低TG,该研究为后续深入揭示CGH蛋白的生理作用及在临床实践中的潜在应用提供了重要基础。

微生物代谢途径调控下的丙酮酸发酵工艺优化与可持续发展

随着当前时代下可持续发展和环境保护意识的不断增强,传统化学合成途径已经无法满足时代发展的要求,而生物发酵作为生产丙酮酸的一种环境友好型方法,受到了相关领域的重点关注。丙酮酸是一种关键的化学中间体,在化工、食品以及医药产业等领域扮演着重要角色。文章主要探讨了微生物代谢途径下的丙酮酸发酵工艺优化与可持续发展的策略及方法,旨在实现丙酮酸的高效、低成本和环境友好的合成。

枯草芽孢杆菌发酵工艺研究进展

综述了枯草芽孢杆菌发酵过程中涉及的多种发酵工程策略,详细探讨了包括高产菌株的选育、培养基的优化、发酵条件的设定以及发酵过程的精细调控等方面的内容。探讨了通过传统育种技术和现代基因工程手段来提高菌株产量的方法。分析了不同营养成分和添加剂对菌株生长和产物生成的影响。介绍了温度、pH、溶氧量等因素对发酵过程的影响及其优化策略。此外,还探讨了发酵过程的动态监控和调控方法,以实现高效的生产过程。对枯草芽孢杆菌工程菌发酵领域的最新研究现状进行了全面介绍,总结了当前的研究成果和存在的问题,提出了未来研究的方向和可能的发展趋势。

植物萜类代谢工程

植物萜类化合物不仅在植物生命活动中起重要作用,而且具有重要商业价值。随着近年来萜类代谢途径和调控机理研究的深入,代谢工程已成为提高萜类产量最有潜力的途径之一。对萜类代谢工程领域具代表性的研究结果进行了全面回顾,然后讨论了萜类代谢工程的研究方法和策略,其中重点探讨了功能基因组学方法在萜类代谢途径及调控机理研究方面的应用。

中药青蒿的生理生化特征及其研究进展

This article presents a brief review concerning the different aspects of studies on Artemisia annua L. Many research papers have been published regarding artificial planting, and physiological and chemical characteristics of A. annua in recent years. The progresses in investigations to enhance artemisinin production from A. annua cell, tissue culture, whole plants, and gene manipulation are discussed in this paper. Fig 2, Ref

植物花香代谢调节与基因工程研究进展

植物花香在吸引昆虫授粉、提高观赏价值和香精的商业价值方面具有重要的作用。随着分子生物学技术的发展,近年来植物花香基因被大量克隆,对花香化合物的合成与代谢的网络调控机制有了更深刻的认识,基因工程改良花香成为可能。对近年来植物花香的合成途径、花香的释放与基因调节、基因工程的研究进展进行了综述,并就存在的问题进行了分析,为花香的分子育种研究提供参考。

农药污染微生物降解研究及应用进展

农药污染与人们的日常生活息息相关,一直是科学家关注的环境污染热点问题。微生物因其种类丰富、分布广泛、适应性强和代谢途径多样的特点显现出自身在农药污染治理方面的优势。本文总结了近10年来,南京农业大学农业部农业环境微生物重点开放实验室在农药残留微生物降解方面取得的进展,从降解性微生物资源的分离收集,到微生物降解代谢途径分析、降解过程中的关键基因或基因簇的克隆,及在微生物遗传操作方法 SEFA PCR(self-formed adaptor PCR)、基因工程菌的构建等方面进行了详细论述,旨在为降解性微生物在环境修复、生物转化等方面的应用提供理论和实践依据。

木质素代谢的生理意义及其遗传控制研究进展

木质素含量及其相关酶系活性与植物的生长发育、抗病性、抗逆性密切相关.在造纸工业中,木质素处理是造成环境污染的重要来源.本文对木质素代谢在植物生长发育过程中的生理意义及近年来通过控制PAL、4CL、CAD、POD等酶的活性调节木质素含量或改变其组分方面的研究进展进行了综合评述,并对今后的林木育种工作进行了展望.

烟草类胡萝卜素代谢的遗传及基因工程研究进展

综述了烟草类胡萝卜素合成和降解途径所涉及的关键基因的分离、功能分析、分子调控及类胡萝卜素代谢的调节和基因工程研究进展,同时对烟草类胡萝卜素代谢的研究方向和应用前景进行了讨论和展望。

第三代基因工程—代谢工程

代谢工程 (MetabolicEngineering) ,亦称途径工程 (PathwayEngineering)和代谢设计 (MetabolicDesign) ,是一门利用分子生物学原理系统分析细胞代谢网络、并通过DNA重组技术合理设计细胞代谢途径及遗传修饰 ,进而完成细胞特性改造的应用性学科。鉴于它涉及到多基因的联合协同表达控制机制 ,与传统的蛋白多肽单基因表达 (第一代基因工程 )以及基因定向突变 (第二代基因工程 )已有显著区别 ,故作者将之看作为第三代基因工程。