Metabolic Engineering Communications最新文献

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Metabolic bottlenecks of Pseudomonas taiwanensis VLB120 during growth on d-xylose via the Weimberg pathway 台湾假单胞菌(Pseudomonas taiwanensis VLB120)通过魏姆伯格途径在二木糖生长过程中的代谢瓶颈
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-06-01 DOI: 10.1016/j.mec.2024.e00241
Philipp Nerke, Jonas Korb, Frederick Haala, Georg Hubmann, Stephan Lütz
{"title":"Metabolic bottlenecks of Pseudomonas taiwanensis VLB120 during growth on d-xylose via the Weimberg pathway","authors":"Philipp Nerke,&nbsp;Jonas Korb,&nbsp;Frederick Haala,&nbsp;Georg Hubmann,&nbsp;Stephan Lütz","doi":"10.1016/j.mec.2024.e00241","DOIUrl":"https://doi.org/10.1016/j.mec.2024.e00241","url":null,"abstract":"<div><p>The microbial production of value-added chemicals from renewable feedstocks is an important step towards a sustainable, bio-based economy. Therefore, microbes need to efficiently utilize lignocellulosic biomass and its dominant constituents, such as <span>d</span>-xylose. <em>Pseudomonas taiwanensis</em> VLB120 assimilates <span>d</span>-xylose via the five-step Weimberg pathway. However, the knowledge about the metabolic constraints of the Weimberg pathway<em>,</em> i.e., its regulation, dynamics, and metabolite fluxes, is limited, which hampers the optimization and implementation of this pathway for bioprocesses. We characterized the Weimberg pathway activity of <em>P. taiwanensis</em> VLB120 in terms of biomass growth and the dynamics of pathway intermediates. In batch cultivations, we found excessive accumulation of the intermediates <span>d</span>-xylonolactone and <span>d</span>-xylonate, indicating bottlenecks in <span>d</span>-xylonolactone hydrolysis and <span>d</span>-xylonate uptake. Moreover, the intermediate accumulation was highly dependent on the concentration of <span>d</span>-xylose and the extracellular pH. To encounter the apparent bottlenecks, we identified and overexpressed two genes coding for putative endogenous xylonolactonases PVLB_05820 and PVLB_12345. Compared to the control strain, the overexpression of PVLB_12345 resulted in an increased growth rate and biomass generation of up to 30 % and 100 %, respectively. Next, <span>d</span>-xylonate accumulation was decreased by overexpressing two newly identified <span>d</span>-xylonate transporter genes, PVLB_18545 and <em>gntP</em> (PVLB_13665). Finally, we combined xylonolactonase overexpression with enhanced uptake of <span>d</span>-xylonate by knocking out the <em>gntP</em> repressor gene <em>gntR</em> (PVLB_13655) and increased the growth rate and biomass yield by 50 % and 24 % in stirred-tank bioreactors, respectively. Our study contributes to the fundamental knowledge of the Weimberg pathway in pseudomonads and demonstrates how to encounter the metabolic bottlenecks of the Weimberg pathway to advance strain developments and cell factory design for bioprocesses on renewable feedstocks.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000105/pdfft?md5=97d4236fe0530871599b5c6105208888&pid=1-s2.0-S2214030124000105-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141314250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bioproduction of methylated phenylpropenes and isoeugenol in Escherichia coli 在大肠杆菌中生物生产甲基化苯丙烯和异丁香酚
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-05-15 DOI: 10.1016/j.mec.2024.e00237
Jeremy Chua, Erik K.R. Hanko, Andrew Yiakoumetti, Ruth A. Stoney, Jakub Chromy, Kris Niño G. Valdehuesa, Katherine A. Hollywood, Cunyu Yan, Eriko Takano, Rainer Breitling
{"title":"Bioproduction of methylated phenylpropenes and isoeugenol in Escherichia coli","authors":"Jeremy Chua,&nbsp;Erik K.R. Hanko,&nbsp;Andrew Yiakoumetti,&nbsp;Ruth A. Stoney,&nbsp;Jakub Chromy,&nbsp;Kris Niño G. Valdehuesa,&nbsp;Katherine A. Hollywood,&nbsp;Cunyu Yan,&nbsp;Eriko Takano,&nbsp;Rainer Breitling","doi":"10.1016/j.mec.2024.e00237","DOIUrl":"https://doi.org/10.1016/j.mec.2024.e00237","url":null,"abstract":"<div><p>Phenylpropenes are a class of natural products that are synthesised by a vast range of plant species and hold considerable promise in the flavour and fragrance industries. Many <em>in vitro</em> studies have been carried out to elucidate and characterise the enzymes responsible for the production of these volatile compounds. However, there is a scarcity of studies demonstrating the <em>in vivo</em> production of phenylpropenes in microbial cell factories. In this study, we engineered <em>Escherichia coli</em> to produce methylchavicol, methyleugenol and isoeugenol from their respective phenylacrylic acid precursors. We achieved this by extending and modifying a previously optimised heterologous pathway for the biosynthesis of chavicol and eugenol. We explored the potential of six <em>S</em>-adenosyl <span>l</span>-methionine (SAM)-dependent <em>O-</em>methyltransferases to produce methylchavicol and methyleugenol from chavicol and eugenol, respectively. Additionally, we examined two isoeugenol synthases for the production of isoeugenol from coniferyl acetate. The best-performing strains in this study were able to achieve titres of 13 mg L<sup>−1</sup> methylchavicol, 59 mg L<sup>−1</sup> methyleugenol and 361 mg L<sup>−1</sup> isoeugenol after feeding with their appropriate phenylacrylic acid substrates. We were able to further increase the methyleugenol titre to 117 mg L<sup>−1</sup> by supplementation with methionine to facilitate SAM recycling. Moreover, we report the biosynthesis of methylchavicol and methyleugenol from <span>l</span>-tyrosine through pathways involving six and eight enzymatic steps, respectively.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000063/pdfft?md5=80f39caf33089f97306dc16312a53f4d&pid=1-s2.0-S2214030124000063-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141067708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Engineering of methionine-auxotroph Escherichia coli via parallel evolution of two enzymes from Corynebacterium glutamicum's direct-sulfurylation pathway enables its recovery in minimal medium 通过并行进化谷氨酸棒状杆菌直接硫化途径中的两种酶来工程化蛋氨酸辅助营养大肠杆菌,使其能够在最小培养基中复原
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-05-10 DOI: 10.1016/j.mec.2024.e00236
Matan Gabay , Inbar Stern , Nadya Gruzdev , Adi Cohen , Lucia Adriana-Lifshits , Tamar Ansbacher , Itamar Yadid , Maayan Gal
{"title":"Engineering of methionine-auxotroph Escherichia coli via parallel evolution of two enzymes from Corynebacterium glutamicum's direct-sulfurylation pathway enables its recovery in minimal medium","authors":"Matan Gabay ,&nbsp;Inbar Stern ,&nbsp;Nadya Gruzdev ,&nbsp;Adi Cohen ,&nbsp;Lucia Adriana-Lifshits ,&nbsp;Tamar Ansbacher ,&nbsp;Itamar Yadid ,&nbsp;Maayan Gal","doi":"10.1016/j.mec.2024.e00236","DOIUrl":"https://doi.org/10.1016/j.mec.2024.e00236","url":null,"abstract":"<div><p>Methionine biosynthesis relies on the sequential catalysis of multiple enzymes. <em>Escherichia coli</em>, the main bacteria used in research and industry for protein production and engineering, utilizes the three-step trans-sulfurylation pathway catalyzed by L-homoserine O-succinyl transferase, cystathionine gamma synthase and cystathionine beta lyase to convert L-homoserine to L-homocysteine. However, most bacteria employ the two-step direct-sulfurylation pathway involving L-homoserine O-acetyltransferases and O-acetyl homoserine sulfhydrylase. We previously showed that a methionine-auxotroph <em>Escherichia</em> <em>coli</em> strain (MG1655) with deletion of <em>metA</em>, encoding for L-homoserine O-succinyl transferase, and <em>metB</em>, encoding for cystathionine gamma synthase, could be complemented by introducing the genes <em>metX</em>, encoding for L-homoserine O-acetyltransferases and <em>metY</em>, encoding for O-acetyl homoserine sulfhydrylase, from various sources, thus altering the <em>Escherichia coli</em> methionine biosynthesis metabolic pathway to direct-sulfurylation. However, introducing <em>metX</em> and <em>metY</em> from <em>Corynebacterium glutamicum</em> failed to complement methionine auxotrophy. Herein, we generated a randomized genetic library based on the <em>metX</em> and <em>metY</em> of <em>Corynebacterium glutamicum</em> and transformed it into a methionine-auxotrophic <em>Escherichia coli</em> strain lacking the <em>metA</em> and <em>metB</em> genes. Through multiple enrichment cycles, we successfully isolated active clones capable of growing in M9 minimal media. The dominant <em>metX</em> mutations in the evolved methionine-autotrophs <em>Escherichia coli</em> were L315P and H46R. Interestingly, we found that a <em>metY</em> gene encoding only the N-terminus 106 out of 438 amino acids of the wild-type MetY enzyme is functional and supports the growth of the methionine auxotroph. Recloning the new genes into the original plasmid and transforming them to methionine auxotroph <em>Escherichia coli</em> validated their functionality. These results show that directed enzyme-evolution enables fast and simultaneous engineering of new active variants within the <em>Escherichia coli</em> methionine direct-sulfurylation pathway, leading to efficient complementation.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000051/pdfft?md5=13216db11f331277ec4bffeddfb976eb&pid=1-s2.0-S2214030124000051-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140946867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Improving 5-(hydroxymethyl)furfural (HMF) tolerance of Pseudomonas taiwanensis VLB120 by automated adaptive laboratory evolution (ALE) 通过实验室自动适应性进化(ALE)提高台湾假单胞菌 VLB120 对 5-(羟甲基)糠醛(HMF)的耐受性
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-05-10 DOI: 10.1016/j.mec.2024.e00235
Thorsten Lechtenberg, Benedikt Wynands, Moritz-Fabian Müller, Tino Polen, Stephan Noack, Nick Wierckx
{"title":"Improving 5-(hydroxymethyl)furfural (HMF) tolerance of Pseudomonas taiwanensis VLB120 by automated adaptive laboratory evolution (ALE)","authors":"Thorsten Lechtenberg,&nbsp;Benedikt Wynands,&nbsp;Moritz-Fabian Müller,&nbsp;Tino Polen,&nbsp;Stephan Noack,&nbsp;Nick Wierckx","doi":"10.1016/j.mec.2024.e00235","DOIUrl":"10.1016/j.mec.2024.e00235","url":null,"abstract":"<div><p>The aldehyde 5-(hydroxymethyl)furfural (HMF) is of great importance for a circular bioeconomy. It is a renewable platform chemical that can be converted into a range of useful compounds to replace petroleum-based products such as the green plastic monomer 2,5-furandicarboxylic acid (FDCA). However, it also exhibits microbial toxicity for example hindering the efficient biotechnological valorization of lignocellulosic hydrolysates. Thus, there is an urgent need for tolerance-improved organisms applicable to whole-cell biocatalysis. Here, we engineer an oxidation-deficient derivative of the naturally robust and emerging biotechnological workhorse <em>P. taiwanensis</em> VLB120 by robotics-assisted adaptive laboratory evolution (ALE). The deletion of HMF-oxidizing enzymes enabled for the first time evolution under constant selection pressure by the aldehyde, yielding strains with consistently improved growth characteristics in presence of the toxicant. Genome sequencing of evolved clones revealed loss-of function mutations in the LysR-type transcriptional regulator-encoding <em>mexT</em> preventing expression of the associated efflux pump <em>mexEF</em>-<em>oprN</em>. This knowledge allowed reverse engineering of strains with enhanced aldehyde tolerance, even in a background of active or overexpressed HMF oxidation machinery, demonstrating a synergistic effect of two distinct tolerance mechanisms.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S221403012400004X/pdfft?md5=d7f841437723f702451889b3caf5d32c&pid=1-s2.0-S221403012400004X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141036491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bottom-up parameterization of enzyme rate constants: Reconciling inconsistent data 酶速率常数的自下而上参数化:调和不一致的数据
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-04-23 DOI: 10.1016/j.mec.2024.e00234
Daniel C. Zielinski , Marta R.A. Matos , James E. de Bree , Kevin Glass , Nikolaus Sonnenschein , Bernhard O. Palsson
{"title":"Bottom-up parameterization of enzyme rate constants: Reconciling inconsistent data","authors":"Daniel C. Zielinski ,&nbsp;Marta R.A. Matos ,&nbsp;James E. de Bree ,&nbsp;Kevin Glass ,&nbsp;Nikolaus Sonnenschein ,&nbsp;Bernhard O. Palsson","doi":"10.1016/j.mec.2024.e00234","DOIUrl":"https://doi.org/10.1016/j.mec.2024.e00234","url":null,"abstract":"<div><p>Kinetic models of metabolism are promising platforms for studying complex metabolic systems and designing production strains. Given the availability of enzyme kinetic data from historical experiments and machine learning estimation tools, a straightforward modeling approach is to assemble kinetic data enzyme by enzyme until a desired scale is reached. However, this type of ‘bottom up’ parameterization of kinetic models has been difficult due to a number of issues including gaps in kinetic parameters, the complexity of enzyme mechanisms, inconsistencies between parameters obtained from different sources, and <em>in vitro-in vivo</em> differences. Here, we present a computational workflow for the robust estimation of kinetic parameters for detailed mass action enzyme models while taking into account parameter uncertainty. The resulting software package, termed MASSef (the Mass Action Stoichiometry Simulation Enzyme Fitting package), can handle standard ‘macroscopic’ kinetic parameters, including K<sub>m</sub>, k<sub>cat</sub>, K<sub>i</sub>, K<sub>eq</sub>, and n<sub>h</sub>, as well as diverse reaction mechanisms defined in terms of mass action reactions and ‘microscopic’ rate constants. We provide three enzyme case studies demonstrating that this approach can identify and reconcile inconsistent data either within <em>in vitro</em> experiments or between <em>in vitro</em> and <em>in vivo</em> enzyme function. We further demonstrate how parameterized enzyme modules can be used to assemble pathway-scale kinetic models consistent with <em>in vivo</em> behavior. This work builds on the legacy of knowledge on kinetic behavior of enzymes by enabling robust parameterization of enzyme kinetic models at scale utilizing the abundance of historical literature data and machine learning parameter estimates.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000038/pdfft?md5=b19129eb61d98f2c6edb816a11548b16&pid=1-s2.0-S2214030124000038-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140817023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
2-Stage microfermentations 2 级微发酵
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-04-08 DOI: 10.1016/j.mec.2024.e00233
Shuai Li , Zhixia Ye , Eirik A. Moreb , Romel Menacho-Melgar , Maximillian Golovsky , Michael D. Lynch
{"title":"2-Stage microfermentations","authors":"Shuai Li ,&nbsp;Zhixia Ye ,&nbsp;Eirik A. Moreb ,&nbsp;Romel Menacho-Melgar ,&nbsp;Maximillian Golovsky ,&nbsp;Michael D. Lynch","doi":"10.1016/j.mec.2024.e00233","DOIUrl":"https://doi.org/10.1016/j.mec.2024.e00233","url":null,"abstract":"<div><p>Cell based factories can be engineered to produce a wide variety of products. Advances in DNA synthesis and genome editing have greatly simplified the design and construction of these factories. It has never been easier to generate hundreds or even thousands of cell factory strain variants for evaluation. These advances have amplified the need for standardized, higher throughput means of evaluating these designs. Toward this goal, we have previously reported the development of engineered <em>E. coli</em> strains and associated 2-stage production processes to simplify and standardize strain engineering, evaluation and scale up. This approach relies on decoupling growth (stage 1), from production, which occurs in stationary phase (stage 2). Phosphate depletion is used as the trigger to stop growth as well as induce heterologous expression. Here, we describe in detail the development of protocols for the evaluation of engineered <em>E. coli</em> strains in 2-stage microfermentations. These protocols are readily adaptable to the evaluation of strains producing a wide variety of protein as well as small molecule products. Additionally, by detailing the approach to protocol development, these methods are also adaptable to additional cellular hosts, as well as other 2-stage processes with various additional triggers.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000026/pdfft?md5=24fb5ce51f4ac60d4daa994c11dcd45e&pid=1-s2.0-S2214030124000026-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140558756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Building blocks needed for mechanistic modeling of bioprocesses: A critical review based on protein production by CHO cells 生物过程机理建模所需的基础模块:基于 CHO 细胞生产蛋白质的重要综述
IF 5.2
Metabolic Engineering Communications Pub Date : 2024-02-29 DOI: 10.1016/j.mec.2024.e00232
Yusmel González-Hernández, Patrick Perré
{"title":"Building blocks needed for mechanistic modeling of bioprocesses: A critical review based on protein production by CHO cells","authors":"Yusmel González-Hernández,&nbsp;Patrick Perré","doi":"10.1016/j.mec.2024.e00232","DOIUrl":"https://doi.org/10.1016/j.mec.2024.e00232","url":null,"abstract":"<div><p>This paper reviews the key building blocks needed to develop a mechanistic model for use as an operational production tool. The Chinese Hamster Ovary (CHO) cell, one of the most widely used hosts for antibody production in the pharmaceutical industry, is considered as a case study. CHO cell metabolism is characterized by two main phases, exponential growth followed by a stationary phase with strong protein production. This process presents an appropriate degree of complexity to outline the modeling strategy. The paper is organized into four main steps: (1) CHO systems and data collection; (2) metabolic analysis; (3) formulation of the mathematical model; and finally, (4) numerical solution, calibration, and validation. The overall approach can build a predictive model of target variables. According to the literature, one of the main current modeling challenges lies in understanding and predicting the spontaneous metabolic shift. Possible candidates for the trigger of the metabolic shift include the concentration of lactate and carbon dioxide. In our opinion, ammonium, which is also an inhibiting product, should be further investigated. Finally, the expected progress in the emerging field of hybrid modeling, which combines the best of mechanistic modeling and machine learning, is presented as a fascinating breakthrough. Note that the modeling strategy discussed here is a general framework that can be applied to any bioprocess.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2024-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030124000014/pdfft?md5=092d00458e357daf7d59391680afef78&pid=1-s2.0-S2214030124000014-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140103570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A new concept of biocatalytic synthesis of acrylic monomers for obtaining water-soluble acrylic heteropolymers 生物催化合成丙烯酸单体以获得水溶性丙烯酸杂聚合物的新概念
IF 5.2
Metabolic Engineering Communications Pub Date : 2023-12-19 DOI: 10.1016/j.mec.2023.e00231
Konstantin V. Lavrov , Anna O. Shemyakina , Elena G. Grechishnikova , Tatyana V. Gerasimova , Tatyana I. Kalinina , Andrey D. Novikov , Tatyana E. Leonova , Ludmila E. Ryabchenko , Telman A. Bayburdov , Alexander S. Yanenko
{"title":"A new concept of biocatalytic synthesis of acrylic monomers for obtaining water-soluble acrylic heteropolymers","authors":"Konstantin V. Lavrov ,&nbsp;Anna O. Shemyakina ,&nbsp;Elena G. Grechishnikova ,&nbsp;Tatyana V. Gerasimova ,&nbsp;Tatyana I. Kalinina ,&nbsp;Andrey D. Novikov ,&nbsp;Tatyana E. Leonova ,&nbsp;Ludmila E. Ryabchenko ,&nbsp;Telman A. Bayburdov ,&nbsp;Alexander S. Yanenko","doi":"10.1016/j.mec.2023.e00231","DOIUrl":"10.1016/j.mec.2023.e00231","url":null,"abstract":"<div><p><em>Rhodococcus</em> strains were designed as model biocatalysts (BCs) for the production of acrylic acid and mixtures of acrylic monomers consisting of acrylamide, acrylic acid, and N-alkylacrylamide (N-isopropylacrylamide). To obtain BC strains, we used, among other approaches, adaptive laboratory evolution (ALE), based on the use of the metabolic pathway of amide utilization. Whole genome sequencing of the strains obtained after ALE, as well as subsequent targeted gene disruption, identified candidate genes for three new amidases that are promising for the development of BCs for the production of acrylic acid from acrylamide. New BCs had two types of amidase activities, acrylamide-hydrolyzing and acrylamide-transferring, and by varying the ratio of these activities in BCs, it is possible to influence the ratio of monomers in the resulting mixtures. Based on these strains, a prototype of a new technological concept for the biocatalytic synthesis of acrylic monomers was developed for the production of water-soluble acrylic heteropolymers containing valuable N-alkylacrylamide units. In addition to the possibility of obtaining mixtures of different compositions, the advantages of the concept are a single starting reagent (acrylamide), more unification of processes (all processes are based on the same type of biocatalyst), and potentially greater safety for personnel and the environment compared to existing chemical technologies.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2023-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030123000147/pdfft?md5=a2807f1fd43527a40cf4152387f2e166&pid=1-s2.0-S2214030123000147-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138989034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Rewiring Saccharomyces cerevisiae metabolism for optimised Taxol® precursors production 重组酿酒酵母代谢优化紫杉醇®前体生产
IF 5.2
Metabolic Engineering Communications Pub Date : 2023-11-15 DOI: 10.1016/j.mec.2023.e00229
Behnaz Nowrouzi , Pablo Torres-Montero , Eduard J. Kerkhoven , José L. Martínez , Leonardo Rios-Solis
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引用次数: 0
Engineering Yarrowia lipolytica for the biosynthesis of geraniol 工程解脂耶氏菌生物合成香叶醇
IF 5.2
Metabolic Engineering Communications Pub Date : 2023-11-02 DOI: 10.1016/j.mec.2023.e00228
Ayushi Agrawal , Zhiliang Yang , Mark Blenner
{"title":"Engineering Yarrowia lipolytica for the biosynthesis of geraniol","authors":"Ayushi Agrawal ,&nbsp;Zhiliang Yang ,&nbsp;Mark Blenner","doi":"10.1016/j.mec.2023.e00228","DOIUrl":"https://doi.org/10.1016/j.mec.2023.e00228","url":null,"abstract":"<div><p>Geraniol is a monoterpene with wide applications in the food, cosmetics, and pharmaceutical industries. Microbial production has largely used model organisms lacking favorable properties for monoterpene production. In this work, we produced geraniol in metabolically engineered <em>Yarrowia lipolytica</em>. First, two plant-derived geraniol synthases (GES) from <em>Catharanthus roseus</em> (Cr) and <em>Valeriana officinalis</em> (Vo) were tested based on previous reports of activity. Both wild type and truncated mutants of GES (without signal peptide targeting chloroplast) were examined by co-expressing with MVA pathway enzymes tHMG1 and IDI1. Truncated CrGES (tCrGES) produced the most geraniol and thus was used for further experimentation. The initial strain was obtained by overexpression of the truncated HMG1, IDI and tCrGES. The acetyl-CoA precursor pool was enhanced by overexpressing mevalonate pathway genes such as ERG10, HMGS or MVK, PMK. The final strain overexpressing 3 copies of tCrGES and single copies of ERG10, HMGS, tHMG1, IDI produced approximately 1 g/L in shake-flask fermentation. This is the first demonstration of geraniol production in <em>Yarrowia lipolytica</em> and the highest de novo titer reported to date in yeast.</p></div>","PeriodicalId":18695,"journal":{"name":"Metabolic Engineering Communications","volume":null,"pages":null},"PeriodicalIF":5.2,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2214030123000111/pdfft?md5=f08729130a21ca40f8b22486294fafb5&pid=1-s2.0-S2214030123000111-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91685392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
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