Engineering and transcriptome study of Saccharomyces cerevisiae to produce ginsenoside compound K by glycerol
Biotechnol J. 2024 Feb;19(2):e2300383. doi: 10.1002/biot.202300383.ABSTRACTSynthetic biology-based engineering of Saccharomyces cerevisiae to produce terpenoid natural products is an effective strategy for their industrial application. Previously, we observed that glycerol addition was beneficial for ginsenoside compound K (CK) production in a S. cerevisiae when it was fermented using the YPD medium. Here, we reconstructed the CK synthesis and glycerol catabolic pathway in a high-yield protopanaxadiol (PPD) S. cerevisiae strain. Remarkably, our engineered strain exhibited the ability to utilize glycerol as the sole carbon ...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Chuanbo Zhang Jinping Tian Jiale Zhang Ruixia Liu Xiaomeng Zhao Wenyu Lu Source Type: research
A refined Uni-vector prime editing system improves genome editing outcomes in mammalian cells
Biotechnol J. 2024 Feb;19(2):e2300353. doi: 10.1002/biot.202300353.ABSTRACTPrime editing is an advanced technology in CRISPR/Cas research with increasing numbers of improved methodologies. The original multi-vector method hampers the efficiency and precision of prime editing and also has inherent difficulty in generating homozygous mutations in mammalian cells. To overcome these technical issues, we developed a Uni-vector prime editing system, wherein the major components for prime editing were constructed in all-in-one plasmids, pPE3-pPuro and pePEmax-pPuro. The Uni-vector prime editing plasmids enhance the editing effici...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Ching-Hui Huang Szu-Ying Chiu Yu-Chi Chou Kou-Juey Wu Source Type: research
A large-scale production of mesenchymal stem cells and their exosomes for an efficient treatment against lung inflammation
Biotechnol J. 2024 Feb;19(2):e2300174. doi: 10.1002/biot.202300174.ABSTRACTMesenchymal stem cells (MSCs) and their produced exosomes have demonstrated inherent capabilities of inflammation-guided targeting and inflammatory modulation, inspiring their potential applications as biologic agents for inflammatory treatments. However, the clinical applications of stem cell therapies are currently restricted by several challenges, and one of them is the mass production of stem cells to satisfy the therapeutic demands in the clinical bench. Herein, a production of human amnion-derived MSCs (hMSCs) at a scale of over 1 × 109 cells...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Jinsong Zhang Ruyi Lin Yingyu Li Jiawen Wang Huiqing Ding Panfeng Fang Yingzhi Huang Jing Shi Jianqing Gao Tianyuan Zhang Source Type: research
Drug-loaded lipid nanoparticles improve the removal rates of the Staphylococcus aureus biofilm
Biotechnol J. 2024 Feb;19(2):e2300159. doi: 10.1002/biot.202300159.ABSTRACTBiofilms of the foodborne pathogen Staphylococcus aureus show improved resistance to antibiotics and are difficult to eliminate. To enhance antibacteria and biofilm dispersion via extracellular matrix diffusion, a new lipid nanoparticle was prepared, which employed a mixture of phospholipids and a 0.8% surfactin shell. In the lipid nanoparticle, 31.56 μg mL-1 of erythromycin was encapsulated. The lipid nanoparticle size was approximately 52 nm and the zeta-potential was -67 mV, which was measured using a Marvin laser particle size analyzer. In addi...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Jiaju Qiao Antuo Hu Haibo Zhou Zhaoxin Lu Fanqiang Meng Changzheng Shi Xiaomei Bie Source Type: research
Improvement of an enzymatic cascade synthesis of nicotinamide mononucleotide via protein engineering and reaction-process reinforcement
Biotechnol J. 2024 Feb;19(2):e2300748. doi: 10.1002/biot.202300748.ABSTRACTEnzymatic synthesis of β-nicotinamide mononucleotide (NMN) from D-ribose has garnered widespread attention due to its cheap material, the use of mild reaction conditions, and the ability to produce highly pure products with the desired optical properties. However, the overall NMN yield of this method is impeded by the low activity of rate-limiting enzymes. The ribose-phosphate diphosphokinase (PRS) and nicotinamide phosphoribosyltransferase (NAMPT), that control the rate of the reaction, were engineered to improve the reaction efficacy. The actives...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Feng Peng Jian Hong Jie Cui Ya-Ni An Qian Guo Qi Shen Feng Cheng Ya-Ping Xue Yu-Guo Zheng Source Type: research
Overexpression of Klotho gene using CRISPR/Cas9 induces apoptosis and inhibits cell motility in the human colorectal cancer cells
This study investigates the role of the KL gene in colorectal cancer by using the CRISPR/Cas9 system to overexpress and knock out (KO) the KL gene in human colorectal cancer cells (Caco-2). The effects of the changes were assessed by gene expression analysis, flow cytometry, scratch wound closure assays, colony formation assays, and immunofluorescence staining. Our results showed that overexpression of the KL gene increased apoptosis and decreased cell motility in cancer cells, whereas knockout of the KL gene had the opposite role. The present study elucidates the mechanisms underlying this role and highlights the potentia...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Merve Nur Soykan Sibel Gunes Source Type: research
Secretory production and characterization of a highly effective chitosanase from Streptomyces coelicolor A3(2) M145 in Pichia pastoris
In this study, a glycoside hydrolase family 46 chitosanase from Streptomyces coelicolor A3(2) M145 was firstly cloned and expressed in Pichia pastoris GS115 (P. pastoris GS115). The recombinant enzyme (CsnA) showed maximal activity at pH 6.0 and 65°C. Both thermal stability and pH stability of CsnA expressed in P. pastoris GS115 were significantly increased compared with homologous expression in Streptomyces coelicolor A3(2). A stable chitosanase activity of 725.7 ± 9.58 U mL-1 was obtained in fed-batch fermentation. It's the highest level of CsnA from Streptomyces coelicolor expressed in P. pastoris so far. The hydrolyt...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Jiayu Liang Shengbin He Jian Sun Haodong Bao Lanyu Cui Source Type: research
Initial cyclic-di-GMP upregulation triggers sporadic cellular expansion leading to improved cellular survival
This study provides us a glimpse of the bacterial behavior of evading potential threats through adapting lifestyle changes via c-di-GMP regulation.PMID:38403404 | DOI:10.1002/biot.202300542 (Source: Biotechnology Journal)
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Yongshuai Yang Siyu Guo Can-Jian Hong Zhao-Xun Liang Chun Loong Ho Source Type: research
Encapsulation of pristine and silica-coated human adipose-derived mesenchymal stem cells in gelatin colloidal hydrogels for tissue engineering and bioprinting applications
Biotechnol J. 2024 Feb;19(2):e2300469. doi: 10.1002/biot.202300469.ABSTRACTColloidal gels assembled from gelatin nanoparticles (GNPs) as particulate building blocks show strong promise to solve challenges in cell delivery and biofabrication, such as low cell survival and limited spatial retention. These gels offer evident advantages to facilitate cell encapsulation, but research on this topic is still limited, which hampers our understanding of the relationship between the physicochemical and biological properties of cell-laden colloidal gels. Human adipose-derived mesenchymal stem cells were successfully encapsulated in g...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Marta M Maciel Negar Hassani Besheli Tiago R Correia Jo ão F Mano Sander C G Leeuwenburgh Source Type: research
Hydrodynamic studies of innovative membrane reactor for enzymatic hydrolysis of lignocellulosic waste
Biotechnol J. 2024 Feb;19(2):e2300602. doi: 10.1002/biot.202300602.ABSTRACTThis paper presents the study concerning the impact of the basic operational parameters on the performance of an innovative microfiltration membrane reactor applied for enzymatic hydrolysis of lignocellulosic biomass. The concept and basic hydrodynamics of the reactor with tubular ceramic membranes and a propeller agitator were shown. Besides, the efficiency of enzymatic hydrolysis of corn straw was studied to check reactor functionality. It has been proven that the proposed reactor construction can improve the microfiltration of lignocellulosic sus...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Katarzyna D ąbkowska-Susfał Joanna Lipi ńska Pawe ł Sobieszuk Andrzej B Ko łtuniewicz Source Type: research
Exploring metabolic responses and pathway changes in CHO-K1 cells under varied aeration conditions and copper supplementations using < sup > 1 < /sup > H NMR-based metabolomics
This study aims to improve our understanding of the CHO-K1 cell metabolism using 1 H NMR-based metabolomics. Initially, the variations in culture performance and metabolic profiles under varied aeration conditions and copper supplementations were thoroughly examined. Furthermore, a comprehensive metabolic pathway analysis was performed to assess the impact of these conditions on the implicated pathways. The results revealed substantial alterations in the pyruvate metabolism, histidine metabolism, as well as phenylalanine, tyrosine and tryptophan biosynthesis, which were especially evident in cultures subjected to copper de...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Yingting Shi Yuxiang Wan Yan Sun Jiayu Yang Yuting Lu Xinyuan Xie Jianyang Pan Haibin Wang Haibin Qu Source Type: research
Synergistic application of atmospheric and room temperature plasma mutagenesis and adaptive laboratory evolution improves the tolerance of Escherichia coli to L-cysteine
In this study, we employed a synergistic approach encompassing atmospheric and room temperature plasma mutagenesis (ARTP) and adaptive laboratory evolution (ALE) to improve L-cysteine tolerance in Escherichia coli. ARTP-treated populations obtained substantial enhancement in L-cysteine tolerance by ALE. Whole-genome sequencing, transcription analysis, and reverse engineering, revealed the pivotal role of an effective export mechanism mediated by gene eamB in augmenting L-cysteine resistance. The isolated tolerant strain, 60AP03/pTrc-cysEf , achieved a 2.2-fold increase in L-cysteine titer by overexpressing the critical gen...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Hui Yang Bo Zhang Zidan Wu Jiayuan Pan Lifeng Chen Xiaoling Xiu Xue Cai Zhiqiang Liu Yuguo Zheng Source Type: research
Exploring engineering strategies that enhance de novo production of exotic cyclopropane fatty acids in Saccharomyces cerevisiae
Biotechnol J. 2024 Feb;19(2):e2300694. doi: 10.1002/biot.202300694.ABSTRACTCycloalkanes have broad applications as specialty fuels, lubricants, and pharmaceuticals but are not currently available from renewable sources, whereas, production of microbial cycloalkanes such as cyclopropane fatty acids (CFA) has bottlenecks. Here, a systematic investigation was undertaken into the biosynthesis of CFA in Saccharomyces cerevisiae heterologously expressing bacterial CFA synthase. The enzyme catalyzes formation of a 3-membered ring in unsaturated fatty acids. Monounsaturated fatty acids in phospholipids (PL) are the site of CFA syn...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Wei Jiang Huadong Peng Lizhong He Rodrigo Lesma-Amaro Victoria S Haritos Source Type: research
The choice of ultra-low attachment plates impacts primary human and primary canine hepatocyte spheroid formation, phenotypes, and function
Biotechnol J. 2024 Feb;19(2):e2300587. doi: 10.1002/biot.202300587.ABSTRACTOrganotypic three-dimensional liver spheroid cultures in which hepatic cells retain their molecular phenotype and functionality have emerged as powerful tools for preclinical drug development. In recent years a multitude of culture systems have been developed; however, a thorough side-by-side benchmarking of the different methods is lacking. Here, we compared the performance of ten different 96- and 384-well microplate types to support spheroid formation and long-term culture. Specifically, we evaluated differences in spheroid formation kinetics, vi...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Chen Xing Aurino Kemas Evgeniya Mickols Kathrin Klein Per Artursson Volker M Lauschke Source Type: research
Cryopreservation of organoids: Strategies, innovation, and future prospects
Biotechnol J. 2024 Feb;19(2):e2300543. doi: 10.1002/biot.202300543.ABSTRACTOrganoid technology has demonstrated unique advantages in multidisciplinary fields such as disease research, tumor drug sensitivity, clinical immunity, drug toxicology, and regenerative medicine. It will become the most promising research tool in translational research. However, the long preparation time of organoids and the lack of high-quality cryopreservation methods limit the further application of organoids. Although the high-quality cryopreservation of small-volume biological samples such as cells and embryos has been successfully achieved, th...
Source: Biotechnology Journal - February 25, 2024 Category: Biotechnology Authors: Hengxin Han Taijie Zhan Ning Guo Mengdong Cui Yi Xu Source Type: research
