Biotechnology and Bioengineering最新文献

{"title":"H2 Consumption by Various Acetogenic Bacteria Follows First‐Order Kinetics up to H2 Saturation","authors":"Laura Muñoz‐Duarte, Susmit Chakraborty, Louise Vinther Grøn, Maria Florencia Bambace, Jacopo Catalano, Jo Philips","doi":"10.1002/bit.28904","DOIUrl":"https://doi.org/10.1002/bit.28904","url":null,"abstract":"Acetogenic bacteria play an important role in various biotechnological processes, because of their chemolithoautotrophic metabolism converting carbon dioxide with molecular hydrogen (H<jats:sub>2</jats:sub>) as electron donor into acetate. As the main factor limiting acetogenesis is often H<jats:sub>2</jats:sub>, insights into the H<jats:sub>2</jats:sub> consumption kinetics of acetogens are required to assess their potential in biotechnological processes. In this study, initial H<jats:sub>2</jats:sub> consumption rates at a range of different initial H<jats:sub>2</jats:sub> concentrations were measured for three different acetogens. Interestingly, for all three strains, H<jats:sub>2</jats:sub> consumption was found to follow first‐order kinetics, i.e. the H<jats:sub>2</jats:sub> consumption rate increased linearly with the dissolved H<jats:sub>2</jats:sub> concentration, up to almost saturated H<jats:sub>2</jats:sub> levels (600 µM). This is in contrast with Monod kinetics and low half‐saturation concentrations, which have commonly been assumed for acetogens. The obtained biomass specific first‐order rate coefficients (<jats:italic>k</jats:italic><jats:sub>1</jats:sub><jats:sup>X</jats:sup>) were further validated by comparison with values obtained by fitting first‐order kinetics on previous time‐course experimental results. The latter method was also used to determine the <jats:italic>k</jats:italic><jats:sub>1</jats:sub><jats:sup>X</jats:sup> value of five additional acetogens strains. Biomass specific first‐order rate coefficients were found to vary up to six‐fold, with the highest <jats:italic>k</jats:italic><jats:sub>1</jats:sub><jats:sup>X</jats:sup> for <jats:italic>Acetobacterium wieringae</jats:italic> and the lowest for <jats:italic>Sporomusa sphaeroides</jats:italic>. Overall, our results demonstrate the importance of the dissolved H<jats:sub>2</jats:sub> concentration to understand the rate of acetogenesis in biotechnological systems.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"48 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142887942","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":"Dual‐Stage Cross‐Flow Filtration: Integrated Capture and Purification of Virus‐Like Particles","authors":"Annabelle Dietrich, Luca Heim, Jürgen Hubbuch","doi":"10.1002/bit.28914","DOIUrl":"https://doi.org/10.1002/bit.28914","url":null,"abstract":"Virus‐like particles (VLPs) are a versatile technology for the targeted delivery of genetic material through packaging and potential surface modifications for directed delivery or immunological issues. Although VLP production is relatively simple as they can be recombinantly produced using microorganisms such as <jats:italic>Escherichia coli</jats:italic>, their current downstream processing often relies on individually developed purification strategies. Integrating size‐selective separation techniques may allow standardized platform processing across VLP purification. This study presents an innovative dual‐stage cross‐flow filtration (CFF) set‐up for integrated capture and purification of VLPs, enabling processing solely based on the size‐selective separation techniques precipitation and filtration. The 2 μm/300 kDa MWCO membrane configuration allows the seamless integration of selective VLP precipitation, two consecutive diafiltration steps–first, for washing the VLP precipitates in the first membrane stage, and second, for isolating the re‐dissolved VLPs by continuously removing precipitant and contaminants in the second membrane stage–and ultrafiltration for setting a target VLP concentration. Compared to a single‐stage CFF set‐up, this dual‐stage CFF set‐up with its integrative, automated design demonstrated the capabilities of product accumulation and contaminant handling while maintaining high productivity. Overall, this study represents a significant advancement toward standardized platform processing of protein nanoparticles through precipitation and filtration, and underscores the potential to expand its applicability to diverse biological molecules, unique process conditions, other phase behavior‐dependent processes, and continuous processing.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"54 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142887046","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":"Recent Research on Chondrocyte Dedifferentiation and Insights for Regenerative Medicine","authors":"Weixian Su, Yupeng Nie, Shicong Zheng, Yongchang Yao","doi":"10.1002/bit.28915","DOIUrl":"https://doi.org/10.1002/bit.28915","url":null,"abstract":"Chondrocytes maintain the balance of the extracellular matrix by synthesizing glycoproteins, collagen, proteoglycans and hyaluronic acid. Chondrocyte dedifferentiation refers to a process in which chondrocytes lose their mature differentiated phenotype and transform into a fibroblast-like morphology with fewer differentiated stages and inferior function under external stimulation. The important mechanism of homeostasis loss in osteoarthritis (OA) is a change in the chondrocyte phenotype. The dedifferentiation markers of chondrocytes are upregulated in OA, and the pathogenic factors related to OA have also been shown to enhance chondrocyte dedifferentiation. In this review, we compile recent studies on chondrocyte dedifferentiation, with an emphasis on potential markers and the underlying mechanisms of dedifferentiation, as well as the current research progress in inhibiting dedifferentiation or achieving redifferentiation. A deep understanding of chondrocyte dedifferentiation would not only support the pathogenesis of OA theoretically but also provide insightful ideas for regenerative medicine to manipulate the functional phenotype of cells.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"11 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142880111","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":"Open-Source Anaerobic Digestion Modeling Platform, Anaerobic Digestion Model No. 1 Fast (ADM1F)","authors":"Kuang Zhu, Wenjuan Zhang, Elchin Jafarov, Satish Karra, Kurt Solander, Meltem Urgun Demirtas, Lutgarde Raskin, Steven Skerlos","doi":"10.1002/bit.28906","DOIUrl":"https://doi.org/10.1002/bit.28906","url":null,"abstract":"An open-source modeling platform, called Anaerobic Digestion Model No. 1 Fast (ADM1F), is introduced to achieve fast and numerically stable simulations of anaerobic digestion processes. ADM1F is compatible with an iPython interface to facilitate model configuration, simulation, data analysis, and visualization. Faster simulations and more stable results are accomplished by implementing an advanced open-source library of numerical methods called Portable Extensive Toolkit for Scientific Computation (PETSc) to solve the ADM1 system of equations. Leveraging PETSc, ADM1F can consistently complete a steady-state simulation under 0.2 s, over 99% faster than a benchmark ADM1 model implemented with MATLAB while achieving agreement of model outputs within 1% of those obtained with the benchmark model. For dynamic simulations, however, ADM1F has a computational speed advantage only when the influent characteristics update more frequently than every 4 h. The ability of ADM1F to be useful as a tool to study anaerobic digestion systems is demonstrated through two example implementations of ADM1F: (1) a two-phase co-digestion scenario evaluating the impact of the organic loading rate and the substrate composition on reactor performance and stability, and (2) a conventional digester scenario assessing the effectiveness of recovery strategies after disruptions that led to instability. These examples demonstrate how the high simulation speed and the convenience of the iPython interface allow ADM1F to complete complex analyses within minutes, much faster than computational strategies currently reported in the literature.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"89 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142880237","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":"Impact of Hormone on Growth and GA3 Regulation of Anthocyanin Biosynthesis in Suspension‐Culture Cells of Cyclocarya paliurus","authors":"Chuanqing Pan, Yuan Liu, Daobang Tang, Jiguang Chen, Zhongping Yin","doi":"10.1002/bit.28913","DOIUrl":"https://doi.org/10.1002/bit.28913","url":null,"abstract":"Hormones are effective in regulating plant metabolism and, therefore, are often used in plant cell culture to increase the yield of target products. This study investigated the effects of hormones on the growth and anthocyanin biosynthesis of suspension‐cultured red <jats:italic>Cyclocarya paliurus</jats:italic> cells. Additionally, the mechanism by which gibberellin induces anthocyanin biosynthesis was explored through multi‐omics integrated analysis and the assay of the dynamic changes in signaling molecule concentration. The results showed that the total anthocyanin content and yield of suspension‐cultured cells, when induced by 1.0 mg L⁻¹ Gibberellin A<jats:sub>3</jats:sub> (GA<jats:sub>3</jats:sub>), experienced increases of 1.92‐ and 1.83‐fold, respectively. The application of exogenous GA<jats:sub>3</jats:sub> activated the synthesis and transduction of four signaling molecules, that is, nitric oxide (NO), hydrogen peroxide (H<jats:sub>2</jats:sub>O<jats:sub>2</jats:sub>), salicylic acid (SA), and jasmonic acid (JA), in the cells and altered the expression patterns of transcription factors. The altered expression of transcription factors upregulated the expression of anthocyanin biosynthetic genes such as <jats:italic>anthocyanin‐3‐O‐glucosyl transferase</jats:italic> and <jats:italic>leucoanthocyanidin dioxygenase</jats:italic>, while downregulated the expression of <jats:italic>anthocyanin reductase</jats:italic> and <jats:italic>flavonoid 3′,5′,‐hydroxylase</jats:italic>, which activated the anthocyanin biosynthesis pathway, ultimately leading to a significant increase in anthocyanin biosynthesis. This research work establishes a foundation for further research on the role of hormones in regulating anthocyanin biosynthesis in suspension‐cultured plant cells.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"78 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2024-12-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142874560","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":"Research on Intelligent Monitoring and Concentration Prediction for Penicillin Fermentation Process","authors":"Yin Zhang,&nbsp;Kai Zhang,&nbsp;Ting Hu,&nbsp;Libo Yuan","doi":"10.1002/bit.28903","DOIUrl":"10.1002/bit.28903","url":null,"abstract":"<div>\u0000 \u0000 <p>In the biopharmaceutical industry, accurately predicting penicillin concentration during fermentation is key to boosting production efficiency and quality assurance. This study leverages the PenSim simulation data set and applies various machine learning and deep learning techniques to forecast penicillin fermentation concentration. Initially, through correlation analysis, nine feature variables with significant impacts on penicillin concentration were screened, and the data underwent preprocessing and standardization. Using grid search, we systematically optimize the hyperparameters of various prediction models. Results show that the ridge regression model excels, achieving a mean squared error of 0.0512 and a mean absolute error of 0.0361. This indicates a strong linear relationship between penicillin concentration and the selected features. Our study offers data-driven insights for intelligent monitoring and optimization of penicillin fermentation processes. It also showcases the potential of artificial intelligence in enhancing control of biotechnological facilities, paving the way for future research.</p>\u0000 </div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 3","pages":"570-578"},"PeriodicalIF":3.5,"publicationDate":"2024-12-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142874561","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":"Efficient Multiplex Genome Editing of the Cyanobacterium Synechocystis sp. PCC6803 via CRISPR-Cas12a","authors":"Wei Du,&nbsp;Luna L. Meister,&nbsp;Tobias van Grinsven,&nbsp;Filipe Branco dos Santos","doi":"10.1002/bit.28910","DOIUrl":"10.1002/bit.28910","url":null,"abstract":"<p>Cyanobacteria have been genetically modified to convert CO₂ into biochemical products, but efficient genetic engineering tools, including CRISPR-Cas systems, remain limited. This is primarily due to the polyploid nature of cyanobacteria, which hinders their effectiveness. Here, we address the latter by specifically (i) modifying the RSF1010-based replicative plasmid to simplify cloning efforts while maintaining high conjugation efficiency; (ii) improving the design of the guide RNA (gRNA) to facilitate chromosomal cleavage; (iii) introducing template DNA fragments as pure plasmids via natural transformation; and (iv) using <i>sacB</i> to facilitate replicative plasmid curing. With this system, the replicative plasmid containing both Cas12a and gRNA is introduced to <i>Synechocystis</i> sp. PCC6803 cells via conjugation to cleave the circular chromosomes. Template DNA plasmid that has meanwhile been assimilated will then repair it achieving the desired genetic modifications. This system was validated by successfully deleting various “neutral” chromosomal loci, both individually and collectively, as well as targeting an essential gene, <i>sll1797</i>. With the <i>sacB</i>-sucrose counter-selection, all deletions were simultaneously made markerless in &lt; 4 weeks. Moreover, we also integrate YFP with various protein degradation tags into the chromosome, allowing for their characterization at the chromosomal level. We foresee this system will greatly facilitate future genome engineering in cyanobacteria.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 3","pages":"736-743"},"PeriodicalIF":3.5,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28910","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857803","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":"Functional Expression and Construction of a Self-Sufficient Cytochrome P450 Chimera for Efficient Steroidal C14α Hydroxylation in Escherichia coli","authors":"Xia Ke,&nbsp;Hong-Duo Dong,&nbsp;Xi-Man Zhao,&nbsp;Xin-Xin Wang,&nbsp;Zhi-Qiang Liu,&nbsp;Yu-Guo Zheng","doi":"10.1002/bit.28911","DOIUrl":"10.1002/bit.28911","url":null,"abstract":"<div>\u0000 \u0000 <p>C14-functionalized steroids enabled diverse biological activities in anti-gonadotropin and anticancer therapy. However, access to C14-functionalized steroids was impeded by the deficiency of chemical synthetic methods. Recently, several membrane-bound fungal cytochrome P450s (CYPs) have been identified with steroid C14α-hydroxylation activity. However, the lack of efficient heterologous overexpression strategy hampered their further characterization and molecular engineering. In the present study, sequences of fungi-derived CYP genes encoding putative 14α-hydroxylase were selected and bioinformatically analyzed. Substitution of the N-terminal hydrophobic helix by a soluble maltose binding protein tag significantly enhanced the soluble expression level in <i>Escherichia coli</i>. A novel CYP originated from <i>Bipolaris oryzae</i> was discovered with high steroidal C14α-hydroxylation activity when coupled with the redox partner CPR_lun. A catalytically self-sufficient chimeric CYP-CPR was built by intramolecular fusion, and the electronic transfer rate was improved. A coenzyme NADPH regeneration system was finally constructed by the co-expression of glucose dehydrogenase. The developed soluble multi-enzyme cascade biotransformation system supported the selective C14α-hydroxylation toward progesterone with a final titer of 34.54 mg/L, the highest level achieved in <i>E. coli</i>-based heterologous expression system. This study provides insightful ideas on the functional expression of fungi-derived CYPs and promises an efficient C14α-hydroxylation system for steroidal drugs through protein engineering.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 3","pages":"724-735"},"PeriodicalIF":3.5,"publicationDate":"2024-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857802","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":"In Situ, High-Resolution Quantification of CO2 Uptake Rates via Automated Off-Gas Analysis Illuminates Carbon Uptake Dynamics in Cyanobacterial Cultures","authors":"Christopher M. Jones,&nbsp;Sean Innes,&nbsp;Steven Holland,&nbsp;Tyson Burch,&nbsp;Sydney Parrish,&nbsp;David R. Nielsen","doi":"10.1002/bit.28905","DOIUrl":"10.1002/bit.28905","url":null,"abstract":"<div>\u0000 \u0000 <p>Quantification of cyanobacterial CO₂ fixation rates is vital to determining their potential as industrial strains in a circular bioeconomy. Currently, however, CO₂ fixation rates are most often determined through indirect and/or low-resolution methods, resulting in an incomplete picture of both dynamic behaviors and total carbon fixation potential. To address this, we developed the “Automated Carbon and CO₂ Experimental Sampling System” (ACCESS); a low-cost system for in situ off-gas analysis that supports the automated acquisition of high-resolution volumetric CO₂ uptake rates from multiple cyanobacterial cultures in parallel. Carbon fixation data obtained via ACCESS were first independently validated by elemental analysis of cultivated biomass. Using ACCESS, we then demonstrate how the volumetric CO₂ uptake rate of two model cyanobacteria, <i>Synechococcus sp</i>. PCC 7002 and <i>Synechocystis sp</i>. PCC 6803, accelerates linearly to a maximum before then decaying monotonically to cessation by stationary phase. Furthermore, consistent with the expected stoichiometry, strong correlations were also found to exist between cell growth and carbon fixation, both in terms of rates as well as total levels. The novel insights made possible via ACCESS will aid other cyanobacterial researchers in diverse fundamental and applied research efforts.</p></div>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 3","pages":"594-605"},"PeriodicalIF":3.5,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142849568","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":"Kinetic Modeling of the Antibody Disulfide Bond Reduction Reaction With Integrated Prediction of the Drug Load Profile for Cysteine-Conjugated ADCs","authors":"Jan Tobias Weggen,&nbsp;Pedro González,&nbsp;Kimberly Hui,&nbsp;Ryan Bean,&nbsp;Michaela Wendeler,&nbsp;Jürgen Hubbuch","doi":"10.1002/bit.28899","DOIUrl":"10.1002/bit.28899","url":null,"abstract":"<p>Antibody-drug conjugates (ADC) constitute a groundbreaking advancement in the field of targeted therapy. In the widely utilized cysteine conjugation, the cytotoxic payload is attached to reduced interchain disulfides which involves a reduction of the native monoclonal antibody (mAb). This reaction needs to be thoroughly understood and controlled as it influences the critical quality attributes (CQAs) of the final ADC product, such as the drug-to-antibody ratio (DAR) and the drug load distribution (DLD). However, existing methodologies lack a mechanistic description of the relationship between process parameters and CQAs. In this context, kinetic modeling provides comprehensive reaction understanding, facilitating the model-based optimization of reduction reaction parameters and potentially reduces the experimental effort needed to develop a robust process. With this study, we introduce an integrated modeling framework consisting of a reduction kinetic model for the species formed during the mAb reduction reaction in combination with a regression model to quantify the number of conjugated drugs by DAR and DLD. The species formed during reduction will be measured by analytical capillary gel electrophoresis (CGE), and the DAR and DLD will be derived from reversed-phase (RP) chromatography. First, we present the development of a reduction kinetic model to describe the impact of reducing agent excess and reaction temperature on the kinetic, by careful investigation of different reaction networks and sets of kinetic rates. Second, we introduce a cross-analytical approach based on multiple linear regression (MLR), wherein CGE data is converted into the RP-derived DAR/DLD. By coupling this with the newly developed reduction kinetic model, an integrated model encompassing the two consecutive reaction steps, reduction and conjugation, is created to predict the final DAR/DLD from initial reduction reaction conditions. The integrated model is finally utilized for an in silico screening to analyze the effect of the reduction conditions, TCEP excess, temperature and reaction time, directly on the final ADC product.</p>","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"122 3","pages":"579-593"},"PeriodicalIF":3.5,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/bit.28899","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142832848","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}