Journal of biotechnology最新文献

{"title":"Recent developments in enzymatic and microbial biosynthesis of flavor and fragrance molecules","authors":"Roman M. Dickey,&nbsp;Madan R. Gopal,&nbsp;Priyanka Nain,&nbsp;Aditya M. Kunjapur","doi":"10.1016/j.jbiotec.2024.04.004","DOIUrl":"10.1016/j.jbiotec.2024.04.004","url":null,"abstract":"<div><p>Flavors and fragrances are an important class of specialty chemicals for which interest in biomanufacturing has risen during recent years. These naturally occurring compounds are often amenable to biosynthesis using purified enzyme catalysts or metabolically engineered microbial cells in fermentation processes. In this review, we provide a brief overview of the categories of molecules that have received the greatest interest, both academically and industrially, by examining scholarly publications as well as patent literature. Overall, we seek to highlight innovations in the key reaction steps and microbial hosts used in flavor and fragrance manufacturing.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140771794","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enzymatic one-step synthesis of natural 2-pyrones and new-to-nature derivatives from coenzyme A esters","authors":"Svitlana Manoilenko ,&nbsp;Martin Dippe ,&nbsp;Tristan Fuchs ,&nbsp;Daniela Eisenschmidt-Bönn ,&nbsp;Jörg Ziegler ,&nbsp;Anne-Katrin Bauer ,&nbsp;Ludger A. Wessjohann","doi":"10.1016/j.jbiotec.2024.04.006","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.04.006","url":null,"abstract":"<div><p>The 2-pyrone moiety is present in a wide range of structurally diverse natural products with various biological activities. The plant biosynthetic routes towards these compounds mainly depend on the activity of either type III polyketide synthase-like 2-pyrone synthases or hydroxylating 2-oxoglutarate dependent dioxygenases. In the present study, the substrate specificity of these enzymes is investigated by a systematic screening using both natural and artificial substrates with the aims of efficiently forming (new) products and understanding the underlying catalytic mechanisms. In this framework, we focused on the <em>in vitro</em> functional characterization of a 2-pyrone synthase Gh2PS2 from <em>Gerbera x hybrida</em> and two dioxygenases AtF6’H1 and AtF6’H2 from <em>Arabidopsis thaliana</em> using a set of twenty aromatic and aliphatic CoA esters as substrates. UHPLC-ESI-HRMSⁿ based analyses of reaction intermediates and products revealed a broad substrate specificity of the enzymes, enabling the facile \"green\" synthesis of this important class of natural products and derivatives in a one-step/one-pot reaction in aqueous environment without the need for halogenated or metal reagents and protective groups. Using protein modeling and substrate docking we identified amino acid residues that seem to be important for the observed product scope.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624001056/pdfft?md5=b70eb2f2513a86c2a36c48dbec3d4ce5&pid=1-s2.0-S0168165624001056-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140645571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ectoine production from a novel bacterial strain and high-purity purification with a cost-effective and single-step method","authors":"Furkan Orhan ,&nbsp;Akın Akıncıoğlu ,&nbsp;Ertuğrul Ceyran","doi":"10.1016/j.jbiotec.2024.04.003","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.04.003","url":null,"abstract":"<div><p>This study marks the exploration into the production of ectoine, a valuable compound with significant potential as an antioxidant, osmoprotectant, anti-inflammatory agent, and stabilizer of cell membranes, proteins, and DNA integrity. Our focus centred on investigating the presence of ectoine and optimizing its production by the novel ectoine producer bacterial strain, <em>Piscibacillus halophilus</em>. For the optimization of ectoine production the effects of carbon and nitrogen sources, salt, pH, agitation and incubation period were optimized by one-factor-at-a-time. We started with an initial ectoine content of 46.92 mg/L, and through a series of optimization processes, we achieved a remarkable increase, resulting in an ectoine content of 1498.2 mg/L. The bacterial species <em>P. halophilus</em> achieved its highest ectoine production after 48 h of incubation, with conditions set at 10 % (w/v) salinity, pH of 7.50, and an agitation speed of 160 rpm. These precise conditions were found to be the most favourable for maximizing ectoine production by this strain. Besides, we have achieved successful purification of ectoine from the crude extract through a streamlined single-step process. This purification method has delivered an exceptional level of purity, surpassing 99.15 %, and an impressive yield of over 99 %. Importantly, we accomplished this using readily available and cost-effective strong acids (HCl) and strong bases (NaOH) to arrange pH gradients. The use of acid and base in the purification process of ectoine reflects an innovative and sustainable methodology.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140604449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Validation of a CFD model for cell culture bioreactors at large scale and its application in scale-up","authors":"Zizhuo Xing ,&nbsp;Gearóid Duane ,&nbsp;Josiah O’Sullivan ,&nbsp;Cynthia Chelius ,&nbsp;Laura Smith ,&nbsp;Michael C. Borys ,&nbsp;Anurag Khetan","doi":"10.1016/j.jbiotec.2024.02.006","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.02.006","url":null,"abstract":"<div><p>Among all the operating parameters that control the cell culture environment inside bioreactors, appropriate mixing and aeration are crucial to ensure sufficient oxygen supply, homogeneous mixing, and CO₂ stripping. A model-based manufacturing facility fit approach was applied to define agitation and bottom air flow rates during the process scale-up from laboratory to manufacturing, of which computational fluid dynamics (CFD) was the core modeling tool. The realizable <em>k</em>-ε turbulent dispersed Eulerian gas-liquid flow model was established and validated using experimental values for the volumetric oxygen transfer coefficient (<em>k</em>_<em>L</em><em>a</em>). Model validation defined the process operating parameter ranges for application of the model, identified mixing issues (e.g., impeller flooding, dissolved oxygen gradients, etc.) and the impact of antifoam on <em>k</em>_<em>L</em><em>a</em>. Using the CFD simulation results as inputs to the models for oxygen demand, gas entrance velocity, and CO₂ stripping aided in the design of the agitation and bottom air flow rates needed to meet cellular oxygen demand, control CO₂ levels, mitigate risks for cell damage due to shear, foaming, as well as fire hazards due to high O₂ levels in the bioreactor gas outlet. The recommended operating conditions led to the completion of five manufacturing runs with a 100% success rate. This model-based approach achieved a seamless scale-up and reduced the required number of at-scale development batches, resulting in cost and time savings of a cell culture commercialization process.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140621138","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adding value to banana farming: Antibody production in post-harvest leaves","authors":"Jasdeep Kaur Darsan Singh ,&nbsp;Purabi Mazumdar ,&nbsp;Rofina Yasmin Othman ,&nbsp;Jennifer Ann Harikrishna","doi":"10.1016/j.jbiotec.2024.04.001","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.04.001","url":null,"abstract":"<div><p>Banana, a globally popular fruit, is widely cultivated in tropical and sub-tropical regions. After fruit harvest, remaining banana plant materials are low-value byproducts, mostly composted or used as fibre or for food packaging. As an aim to potentially increase farmer income, this study explored underutilised banana biomass as a novel plant tissue for production of a high-value product. Protein scFvTG130 used in this study, is an anti-toxoplasma single chain variable fragment antibody that can be used in diagnostics and neutralising the <em>Toxoplasma gondii</em> pathogen. Using detached banana leaves, we investigated the factors influencing the efficacy of a transient expression system using reporter genes and recombinant protein, scFvTG130. Transient expression was optimal at 2 days after detached banana leaves were vacuum infiltrated at 0.08 MPa vacuum pressure for a duration of 3 min with 0.01% (v/v) Tween20 using <em>Agrobacterium</em> strain GV3101 harbouring disarmed virus-based vector pIR-GFPscFv<em>TG130</em>. The highest concentration of anti-toxoplasma scFvTG130 antibody obtained using detached banana leaves was 22.8 µg/g fresh leaf tissue. This first study using detached banana leaf tissue for the transient expression of a recombinant protein, successfully demonstrated anti-toxoplasma scFvTG130 antibody expression, supporting the potential application for other related proteins using an underutilised detached banana leaf tissue.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140548992","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Purification, functional characterization and enhanced production of serratiopeptidase from Serratia marcescens MES-4: An endophyte isolated from Morus rubra","authors":"Diksha Koul ,&nbsp;Devtulya Chander ,&nbsp;Ravi S. Manhas ,&nbsp;Md. Mehedi Hossain ,&nbsp;Mohd Jamal Dar ,&nbsp;Asha Chaubey","doi":"10.1016/j.jbiotec.2024.04.002","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.04.002","url":null,"abstract":"<div><p>Serratiopeptidase, a proteolytic enzyme serves as an important anti-inflammatory and analgesic medication. Present study reports the production and purification of extracellular serratiopeptidase from an endophyte, <em>Serratia marcescens</em> MES-4, isolated from <em>Morus rubra</em>. Purification of the enzyme by Ion exchange chromatography led to the specific activity of 13,030 U/mg protein of serratiopeptidase, showcasing about 3.1 fold enhanced activity. The catalytic domain of the purified serratiopeptidase, composed of Zn coordinated with three histidine residues (His 209, His 213, and His 219), along with glutamate (Glu 210) and tyrosine (Tyr 249). The molecular mass, as determined by SDS-PAGE was ∼51 kDa. The purified serratiopeptidase displayed optimal activity at pH 9.0, temperature 50°C. Kinetic studies revealed V_max and K_m values of 33,333 U/mL and 1.66 mg/mL, respectively. Further, optimized conditions for the production of serratiopeptidase by Taguchi design led to the productivity of 87 U/mL/h with 87.9 fold enhanced production as compared to the previous conditions.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140539776","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reduction of anthranilic acid to 2-aminobenzaldehyde by the white-rot fungus Bjerkandera adusta DSMZ 4708","authors":"Valeriia Babkina ,&nbsp;Yana Haiduk ,&nbsp;Yuliia Kurtash ,&nbsp;Holger Zorn ,&nbsp;Tatyana Zhuk","doi":"10.1016/j.jbiotec.2024.03.015","DOIUrl":"https://doi.org/10.1016/j.jbiotec.2024.03.015","url":null,"abstract":"<div><p>The biocatalytic aerobic “in-water” reduction of anthranilic acid to 2-aminobenzaldehyde by growing cultures of the basidiomycetous white-rot fungus <em>Bjerkandera adusta</em> has been studied. The high specific activity of <em>Bjerkandera adusta</em> towards the carboxylic group of anthranilic acid that allows avoiding the formation of the corresponding alcohol has been demonstrated using different substrate concentrations. The presence of ethanol as co-solvent allows increasing the yield of target product. In contrast to chemical reducing agents that usually yield 2-aminobenzyl alcohol, an overreduction of anthranilic acid is completely suppressed by the fungus and gives the target flavor compound in satisfactory preparative yields. It was shown that the activity of <em>Bjerkandera adusta</em> towards anthranilic acid does not apply to its <em>m-</em> and <em>p-</em>isomers.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624000919/pdfft?md5=e183fab869ca32efc32b238649a2ee90&pid=1-s2.0-S0168165624000919-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140539775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing the soluble expression of α-1,2-fucosyltransferase in E. coli using high-throughput flow cytometry screening coupled with a split-GFP","authors":"Jun-Min Lee ,&nbsp;Jung Hwa Kim ,&nbsp;Jin Young Kim ,&nbsp;Min-Kyu Oh ,&nbsp;Byung-Gee Kim","doi":"10.1016/j.jbiotec.2024.03.014","DOIUrl":"10.1016/j.jbiotec.2024.03.014","url":null,"abstract":"<div><p>2’-Fucosyllactose (2’-FL), one of the major human milk oligosaccharides, was produced in several engineered microorganisms. However, the low solubility of α-1,2-fucosyltransferase (α1,2-FucT) often becomes a bottleneck to produce maximum amount of 2’-FL in the microorganisms. To overcome this solubility issue, the following studies were conducted to improve the soluble expression of α1,2-FucT. Initially, hydrophobic amino acids in the hydrophilic region of the 6 α-helices were mutated, adhering to the α-helix rule. Subsequently, <em>gfp</em>11 was fused to the C-terminal of <em>futC</em> gene encoding α1,2-FucT (FutC), enabling selection of high-fluorescence mutants through split-GFP. Each mutant library was screened via fluorescence activated cell sorting (FACS) to separate soluble mutants for high-throughput screening. As a result, L80C single mutant and A121D/P124A/L125R triple mutant were found, and a combined quadruple mutant was created. Furthermore, we combined mutations of conserved sequences (Q150H/C151R/Q239S) of FutC, which showed positive effects in the previous studies from our lab, with the above quadruple mutants (L80C/A121D/P124A/L125R). The resulting strain produced approximately 3.4-fold higher 2′-FL titer than that of the wild-type, suggesting that the conserved sequence mutations are an independent subset of the mutations that further improve the solubility of the target protein acquired by random mutagenesis using split-GFP.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140331749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modification of the endoplasmic reticulum morphology enables improved recombinant antibody expression in Saccharomyces cerevisiae","authors":"Laura R.K. Niemelä,&nbsp;Essi V. Koskela,&nbsp;Alexander D. Frey","doi":"10.1016/j.jbiotec.2024.03.009","DOIUrl":"10.1016/j.jbiotec.2024.03.009","url":null,"abstract":"<div><p>The yeast <em>Saccharomyces cerevisiae</em> is a versatile cell factory used for manufacturing of a wide range of products, among them recombinant proteins. Protein folding is one of the rate-limiting processes and this shortcoming is often overcome by the expression of folding catalysts and chaperones in the endoplasmic reticulum (ER). In this work, we aimed to establish the impact of ER structure on cellular productivity. The reticulon proteins Rtn1p and Rtn2p, and Yop1p are membrane curvature inducing proteins that define the morphology of the ER and depletion of these proteins creates yeast cells with a higher ER sheet-to-tubule ratio. We created yeast strains with different combinations of deletions of Rtn1p, Rtn2p, and Yop1p coding genes in cells with a normal or expanded ER lumen. We identified strains that reached up to 2.2-fold higher antibody titres compared to the control strain. The expanded ER membrane reached by deletion of the lipid biosynthesis repressor <em>OPI1</em> was essential for the increased productivity. The improved specific productivity was accompanied by an up to 2-fold enlarged ER surface area and a 1.5-fold increased cross-sectional cell area. Furthermore, the strains with enlarged ER displayed an attenuated unfolded protein response. These results underline the impact that ER structures have on productivity and support the notion that reprogramming subcellular structures belongs into the toolbox of synthetic biology.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0168165624000804/pdfft?md5=0bb446dee6daea2507f28d0019ca4687&pid=1-s2.0-S0168165624000804-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140329830","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Practical deployment of automation to expedite aqueous two-phase extraction","authors":"Mario A. Torres-Acosta ,&nbsp;Alex Olivares-Molina ,&nbsp;Ross Kent ,&nbsp;Nuno Leitão ,&nbsp;Markus Gershater ,&nbsp;Brenda Parker ,&nbsp;Gary J. Lye ,&nbsp;Duygu Dikicioglu","doi":"10.1016/j.jbiotec.2024.03.013","DOIUrl":"10.1016/j.jbiotec.2024.03.013","url":null,"abstract":"<div><p>The feasibility of bioprocess development relies heavily on the successful application of primary recovery and purification techniques. Aqueous two-phase extraction (ATPE) disrupts the definition of \"unit operation\" by serving as an integrative and intensive technique that combines different objectives such as the removal of biomass and integrated recovery and purification of the product of interest. The relative simplicity of processing large samples renders this technique an attractive alternative for industrial bioprocessing applications. However, process development is hindered by the lack of easily predictable partition behaviours, the elucidation of which necessitates a large number of experiments to be conducted. Liquid handling devices can assist to address this problem; however, they are configured to operate using low viscosity fluids such as water and water-based solutions as opposed to highly viscous polymeric solutions, which are typically required in ATPE. In this work, an automated high throughput ATPE process development framework is presented by constructing phase diagrams and identifying the binodal curves for PEG6000, PEG3000, and PEG2000. Models were built to determine viscosity- and volume-independent transfer parameters. The framework provided an appropriate strategy to develop a very precise and accurate operation by exploiting the relationship between different liquid transfer parameters and process error. Process accuracy, measured by mean absolute error, and device precision, evaluated by the coefficient of variation, were both shown to be affected by the mechanical properties, particularly viscosity, of the fluids employed. For PEG6000, the mean absolute error improved by six-fold (from 4.82% to 0.75%) and the coefficient of variation improved by three-fold (from 0.027 to 0.008) upon optimisation of the liquid transfer parameters accounting for the viscosity effect on the PEG-salt buffer utilising ATPE operations. As demonstrated here, automated liquid handling devices can serve to streamline process development for APTE enabling wide adoption of this technique in large scale bioprocess applications.</p></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":null,"pages":null},"PeriodicalIF":4.1,"publicationDate":"2024-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S016816562400083X/pdfft?md5=14db61af915d8cfde8aabac2a9780df3&pid=1-s2.0-S016816562400083X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140329831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}