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Medium Chain Length Polyhydroxyalkanoate Production by Engineered Pseudomonas gessardii Using Acetate-formate as Carbon Sources
Woo Young Kim, Seung-Jin Kim, Hye-Rin Seo, Yoonyong Yang, Jong Seok Lee, Moonsuk Hur, Byoung-Hee Lee, Jong-Geol Kim, Min-Kyu Oh
J. Microbiol. 2024;62(7):569-579.   Published online May 3, 2024
DOI: https://doi.org/10.1007/s12275-024-00136-x
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AbstractAbstract
Production of medium chain length polyhydroxyalkanoate (mcl-PHA) was attempted using Pseudomonas gessardii NIBRBAC000509957, which was isolated from Sunchang, Jeollabuk-do, Republic of Korea (35°24'27.7"N, 127°09'13.0"E) and effectively utilized acetate and formate as carbon sources. We first evaluated the utilization of acetate as a carbon source, revealing optimal growth at 5 g/L acetate. Then, formate was supplied to the acetate minimal medium as a carbon source to enhance cell growth. After overexpressing the acetate and formate assimilation pathway enzymes, this strain grew at a significantly higher rate in the medium. As this strain naturally produces PHA, it was further engineered metabolically to enhance mcl-PHA production. The engineered strain produced 0.40 g/L of mcl-PHA with a biomass content of 30.43% in fed-batch fermentation. Overall, this strain can be further developed to convert acetate and formate into valuable products.
Enhancement of the solubility of recombinant proteins by fusion with a short-disordered peptide
Jun Ren , Suhee Hwang , Junhao Shen , Hyeongwoo Kim , Hyunjoo Kim , Jieun Kim , Soyoung Ahn , Min-gyun Kim , Seung Ho Lee , Dokyun Na
J. Microbiol. 2022;60(9):960-967.   Published online July 14, 2022
DOI: https://doi.org/10.1007/s12275-022-2122-z
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  • 5 Web of Science
  • 5 Crossref
AbstractAbstract
In protein biotechnology, large soluble fusion partners are widely utilized for increased yield and solubility of recombinant proteins. However, the production of additional large fusion partners poses an additional burden to the host, leading to a decreased protein yield. In this study, we identified two highly disordered short peptides that were able to increase the solubility of an artificially engineered aggregationprone protein, GFP-GFIL4, from 0.6% to 61% (D3-DP00592) and 46% (D4-DP01038) selected from DisProt database. For further confirmation, the peptides were applied to two insoluble E. coli proteins (YagA and YdiU). The peptides also enhanced solubility from 52% to 90% (YagA) and from 27% to 93% (YdiU). Their ability to solubilize recombinant proteins was comparable with strong solubilizing tags, maltosebinding protein (40 kDa) and TrxA (12 kDa), but much smaller (< 7 kDa) in size. For practical application, the two peptides were fused with a restriction enzyme, I-SceI, and they increased I-SceI solubility from 24% up to 75%. The highly disordered peptides did not affect the activity of I-SceI while I-SceI fused with MBP or TrxA displayed no restriction activity. Despite the small size, the highly disordered peptides were able to solubilize recombinant proteins as efficiently as conventional fusion tags and did not interfere with the function of recombinant proteins. Consequently, the identified two highly disordered peptides would have practical utility in protein biotechnology and industry.

Citations

Citations to this article as recorded by  
  • Synthetic intrinsically disordered protein fusion tags that enhance protein solubility
    Nicholas C. Tang, Jonathan C. Su, Yulia Shmidov, Garrett Kelly, Sonal Deshpande, Parul Sirohi, Nikhil Peterson, Ashutosh Chilkoti
    Nature Communications.2024;[Epub]     CrossRef
  • Biosynthesis of Indigo Dyes and Their Application in Green Chemical and Visual Biosensing for Heavy Metals
    Yan Guo, Shun-Yu Hu, Can Wu, Chao-Xian Gao, Chang-Ye Hui
    ACS Omega.2024; 9(31): 33868.     CrossRef
  • Functional small peptides for enhanced protein delivery, solubility, and secretion in microbial biotechnology
    Hyang-Mi Lee, Thi Duc Thai, Wonseop Lim, Jun Ren, Dokyun Na
    Journal of Biotechnology.2023; 375: 40.     CrossRef
  • Directed Evolution of Soluble α-1,2-Fucosyltransferase Using Kanamycin Resistance Protein as a Phenotypic Reporter for Efficient Production of 2'-Fucosyllactose
    Jonghyeok Shin, Seungjoo Kim, Wonbeom Park, Kyoung Chan Jin, Sun-Ki Kim, Dae-Hyuk Kweon
    Journal of Microbiology and Biotechnology.2022; 32(11): 1471.     CrossRef
  • Effects of spray drying, freeze drying, and vacuum drying on physicochemical and nutritional properties of protein peptide powder from salted duck egg white
    Tianyin Du, Jicheng Xu, Shengnan Zhu, Xinjun Yao, Jun Guo, Weiqiao Lv
    Frontiers in Nutrition.2022;[Epub]     CrossRef
Cr(VI) removal from aqueous solution by thermophilic denitrifying bacterium Chelatococcus daeguensis TAD1 in the presence of single and multiple heavy metals
Han Li , Shaobin Huang , Yongqing Zhang
J. Microbiol. 2016;54(9):602-610.   Published online August 31, 2016
DOI: https://doi.org/10.1007/s12275-016-5295-5
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  • 13 Crossref
AbstractAbstract
Cr(VI) pollution is increasing continuously as a result of ongoing industrialization. In this study, we investigated the thermophilic denitrifying bacterium Chelatococcus daeguensis TAD1, isolated from the biofilm of a biotrickling filter used in nitrogen oxides (NOX) removal, with respect to its ability to remove Cr(VI) from an aqueous solution. TAD1 was capable of reducing Cr(VI) from an initial concentration of 10 mg/L to non-detectable levels over a pH range of 7–9 and at a temperature range of 30–50°C. TAD1 simultaneously removed both Cr(VI) and NO3 −-N at 50°C, when the pH was 7 and the initial Cr(VI) concentration was 15 mg/L. The reduction of Cr(VI) to Cr(III) correlated with the growth metabolic activity of TAD1. The presence of other heavy metals (Cu, Zn, and Ni) inhibited the ability of TAD1 to remove Cr(VI). The metals each individually inhibited Cr(VI) removal, and the extent of inhibition increased in a cooperative manner in the presence of a combination of the metals. The addition of biodegradable cellulose acetate microspheres (an adsorption material) weakened the toxicity of the heavy metals; in their presence, the Cr(VI) removal efficiency returned to a high level. The feasibility and applicability of simultaneous nitrate removal and Cr(VI) reduction by strain TAD1 is promising, and may be an effective biological method for the clean-up of wastewater.

Citations

Citations to this article as recorded by  
  • Hexavalent Chromium Pollution and its Sustainable Management through Bioremediation
    Anushka Paul, Sudeshna Dey, Deo Karan Ram, Alok Prasad Das
    Geomicrobiology Journal.2024; 41(4): 324.     CrossRef
  • Adsorption of Hg2+/Cr6+ by metal-binding proteins heterologously expressed in Escherichia coli
    Shuting Hu, Zixiang Wei, Teng Liu, Xinyu Zuo, Xiaoqiang Jia
    BMC Biotechnology.2024;[Epub]     CrossRef
  • Growth-dependent cr(VI) reduction by Alteromonas sp. ORB2 under haloalkaline conditions: toxicity, removal mechanism and effect of heavy metals
    G. Kiran Kumar Reddy, K. Kavibharathi, Anuroop Singh, Y. V. Nancharaiah
    World Journal of Microbiology and Biotechnology.2024;[Epub]     CrossRef
  • Hexavalent Cr, Its Toxicity and Removal Strategy: Revealing PGPB Potential in Its Remediation
    Akanksha Gupta, Anubhuti Singh, Virendra Kumar Mishra
    Water, Air, & Soil Pollution.2023;[Epub]     CrossRef
  • Chromium Toxicity in Plants: Signaling, Mitigation, and Future Perspectives
    Sajad Ali, Rakeeb A. Mir, Anshika Tyagi, Nazia Manzar, Abhijeet Shankar Kashyap, Muntazir Mushtaq, Aamir Raina, Suvin Park, Sandhya Sharma, Zahoor A. Mir, Showkat A. Lone, Ajaz A. Bhat, Uqab Baba, Henda Mahmoudi, Hanhong Bae
    Plants.2023; 12(7): 1502.     CrossRef
  • A comprehensive review on chromium (Cr) contamination and Cr(VI)-resistant extremophiles in diverse extreme environments
    Zeeshanur Rahman, Lebin Thomas, Siva P. K. Chetri, Shrey Bodhankar, Vikas Kumar, Ravi Naidu
    Environmental Science and Pollution Research.2023; 30(21): 59163.     CrossRef
  • Health hazards of hexavalent chromium (Cr (VI)) and its microbial reduction
    Pooja Sharma, Surendra Pratap Singh, Sheetal Kishor Parakh, Yen Wah Tong
    Bioengineered.2022; 13(3): 4923.     CrossRef
  • Reduced graphene oxide supported nanoscale zero-valent iron (nZVI/rGO) for in-situ remediation of Cr(VI)/nitrate-polluted aquifer
    Xinyang Liu, Wanting Liu, Zifang Chi
    Journal of Water Process Engineering.2022; 49: 103188.     CrossRef
  • Simultaneous denitrification and hexavalent chromium removal by a newly isolated Stenotrophomonas maltophilia strain W26 under aerobic conditions
    Qiang An, Shu-man Deng, Bin Zhao, Zheng Li, Jia Xu, Jia-Li Song
    Environmental Chemistry.2021; 18(1): 20.     CrossRef
  • Leucobacter coleopterorum sp. nov., Leucobacter insecticola sp. nov., and Leucobacter viscericola sp. nov., isolated from the intestine of the diving beetles, Cybister brevis and Cybister lewisianus, and emended description of the genus Leucobacter
    Dong-Wook Hyun, Hojun Sung, Pil Soo Kim, Ji-Hyun Yun, Jin-Woo Bae
    Journal of Microbiology.2021; 59(4): 360.     CrossRef
  • Bioremediation of Chromium by Microorganisms and Its Mechanisms Related to Functional Groups
    Abate Ayele, Yakob Godebo Godeto, Yifeng Zhang
    Journal of Chemistry.2021; 2021: 1.     CrossRef
  • Iron oxide minerals promote simultaneous bio-reduction of Cr(VI) and nitrate: Implications for understanding natural attenuation
    Yutian Hu, Tong Liu, Nan Chen, Chuanping Feng
    Science of The Total Environment.2021; 786: 147396.     CrossRef
  • Cr(VI) reductase activity locates in the cytoplasm of Aeribacillus pallidus BK1, a novel Cr(VI)-reducing thermophile isolated from Tengchong geothermal region, China
    Yan Ma, Hui Zhong, Zhiguo He
    Chemical Engineering Journal.2019; 371: 524.     CrossRef
High Efficiency Transformation by Electroporation of Yarrowia lipolytica
Jia-Hung Wang , Wenpin Hung , Shu-Hsien Tsai
J. Microbiol. 2011;49(3):469-472.   Published online June 30, 2011
DOI: https://doi.org/10.1007/s12275-011-0433-6
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AbstractAbstract
Yarrowia lipolytica was usually transformed by heat shock, but linearized integrative vectors always resulted in a low transformation efficiency when electroporation was used. To develop a high efficiency integrative transformation method by electroporation of Y. lipolytica, we report here that pretreatment of Y. lipolytica with 150 mM LiAc for 1 h before electroporation will approximately 30-fold of increase transformation efficiency. A cell concentration of 1010/ml and instrument settings of 1.5 kV will generate the highest transformation efficiencies. We have developed a procedure to transform Y. lipolytica that will be able to yield an efficiency of 2.1×104 transformants/μg for integrative linear DNA. With our modifications, the electroporation procedures became a very efficient and reliable tool for Y. lipolytica transformation.
Research Support, Non-U.S. Gov't
The Schizosaccharomyces pombe Gene Encoding [gamma]-Glutamyl Transpeptidase I Is Regulated by Non-fermentable Carbon Sources and Nitrogen Starvation
Hong-Gyum Kim , Hey-Jung Park , Hyun-Jung Kang , Hye-Won Lim , Kyunghoon Kim , Eun-Hee Park , Kisup Ahn , Chang-Jin Lim
J. Microbiol. 2005;43(1):44-48.
DOI: https://doi.org/2139 [pii]
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AbstractAbstract
In our previous study, the first structural gene (GGTI) encoding g-glutamyl transpeptidase was cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its transcription, using the GGTI-lacZ fusion gene, containing the 1,085 bp upstream region from the translational initiation point, was found to be enhanced by sodium nitroprusside and L-buthionine-(S,R)-sulfoximine (BSO). In the present work, regulation of the GGTI gene was further elucidated. Non-fermentable carbon sources, such as acetate and ethanol, markedly enhanced the synthesis of [beta]-galactosidase from the GGTI-lacZ fusion gene. However, its induction by non-fermentable carbon sources appeared to be independent of the presence of the Pap1 protein. Nitrogen starvation also gave rise to induction of GGTI gene expression in a Pap1-independent manner. The three additional fusion plasmids, carrying 754, 421 and 156 bp regions, were constructed. The sequence responsible for the induction by non-fermentable carbon sources and nitrogen starvation was identified to exist within a -421 bp region of the GGTI gene. Taken together, the S. pombe GGTI gene is regulated by non-fermentable carbon sources and nitrogen starvation.
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