Constitutive Optimized Production of Streptokinase in<i>Saccharomyces cerevisiae</i>Utilizing Glyceraldehyde 3-Phosphate Dehydrogenase Promoter of<i>Pichia pastoris</i>

Vellanki, Ravi N.; Potumarthi, Ravichandra; Doddapaneni, Kiran Kumar; Anubrolu, Naveen; Mangamoori, Lakshmi Narasu

doi:10.1155/2013/268249

Cited by 15 publications

(4 citation statements)

References 32 publications

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“…Due to the size disparities between the protein components, gel filtration can be used to separate them (Mahmoud et al, 2020). The resolved protein is then passed through an absorbent bed and an uncharged gel to recover it (Vellanki et al, 2013). The purification results are demonstrated in Figure 6 present below.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of clinical efficacy of streptokinase by comparison with the thrombolytic agent on animal model

Yousaf,

Arshad,

Harraz

et al. 2024

Braz. J. Biol.

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Cardiovascular disorders, including acute myocardial infarction (AMI), often lead to blood clot formation, impacting blood circulation. Streptokinase, a cost-effective and widely available thrombolytic agent, is crucial in treating thrombosis. This study aimed to produce streptokinase from Streptococcus pyogenes EBL-48 and compare its efficacy with heparin in an animal model. We evaluated the clot-lysing effectiveness of streptokinase produced from Streptococcus pyogenes EBL-48, emphasizing its low cost and ease of production. Streptokinase was produced using pre-optimized fermentation media and purified through ion exchange and gel-filtration chromatography. In vivo analysis involved inducing clots in a trial animal model using ferric chloride, comparing streptokinase with heparin. Ultrasonography assessed the clot-lysing activity of streptokinase. Streptokinase (47 kDa) effectively lysed clots, proving its low cost, easy production, and minimal adverse effects. Ultrasonography confirmed its fibrinolytic efficacy. These findings highlight potential as an affordable and easily produced thrombolytic agent, particularly relevant in resource-limited settings. Streptokinase efficacy and minimal adverse effects make it a promising option for thrombolytic therapy, especially in economically constrained regions. Future studies could optimize production techniques, explore different strains, and conduct clinical trials for human validation. Comparative studies with other thrombolytic agents would enhance understanding of their advantages and limitations.

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Section: Discussionmentioning

confidence: 99%

Evaluation of clinical efficacy of streptokinase by comparison with the thrombolytic agent on animal model

Yousaf,

Arshad,

Harraz

et al. 2024

Braz. J. Biol.

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“…Yeast extract was still used in the optimized medium due to its role in enhancing 2,3-BD titer. Feasibility of using other inexpensive nitrogen source such as corn steep liquor [ 63 , 64 ] to partially substitute yeast extract could be explored with similar medium optimization strategy described in this work.…”

Section: Discussionmentioning

confidence: 99%

Production of (2R, 3R)-2,3-butanediol using engineered Pichia pastoris: strain construction, characterization and fermentation

Yang

Zhang

2018

Biotechnol Biofuels

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Background2,3-butanediol (2,3-BD) is a bulk platform chemical with various potential applications such as aviation fuel. 2,3-BD has three optical isomers: (2R, 3R)-, (2S, 3S)- and meso-2,3-BD. Optically pure 2,3-BD is a crucial precursor for the chiral synthesis and it can also be used as anti-freeze agent due to its low freezing point. 2,3-BD has been produced in both native and non-native hosts. Several pathogenic bacteria were reported to produce 2,3-BD in mixture of its optical isomers including Klebsiella pneumoniae and Klebsiella oxytoca. Engineered hosts based on episomal plasmid expression such as Escherichia coli, Saccharomyces cerevisiae and Bacillus subtilis are not ideal for industrial fermentation due to plasmid instability.ResultsPichia pastoris is generally regarded as safe and a well-established host for high-level heterologous protein production. To produce pure (2R, 3R)-2,3-BD enantiomer, we developed a P. pastoris strain by introducing a synthetic pathway. The alsS and alsD genes from B. subtilis were codon-optimized and synthesized. The BDH1 gene from S. cerevisiae was cloned. These three pathway genes were integrated into the genome of P. pastoris and expressed under the control of GAP promoter. Production of (2R, 3R)-2,3-BD was achieved using glucose as feedstock. The optical purity of (2R, 3R)-2,3-BD was more than 99%. The titer of (2R, 3R)-2,3-BD reached 12 g/L with 40 g/L glucose as carbon source in shake flask fermentation. The fermentation conditions including pH, agitation speeds and aeration rates were optimized in batch cultivations. The highest titer of (2R, 3R)-2,3-BD achieved in fed-batch fermentation using YPD media was 45 g/L. The titer of 2,3-BD was enhanced to 74.5 g/L through statistical medium optimization.ConclusionsThe potential of engineering P. pastoris into a microbial cell factory for biofuel production was evaluated in this work using (2R, 3R)-2,3-BD as an example. Engineered P. pastoris could be a promising workhorse for the production of optically pure (2R, 3R)-2,3-BD.Electronic supplementary materialThe online version of this article (10.1186/s13068-018-1031-1) contains supplementary material, which is available to authorized users.

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“…USM-PSY62 based on 18S rRNA (Ong, 2006). USM-PSY62 was cultured and preserved in YPD medium (1% yeast extract, 2% polypeptone and 2% dextrose) at 15 °C (Vellanki, 2013) and stored for long-term use by preparing a cell suspension in YPD supplemented with 20% glycerol and frozen at -80 °C.…”

Section: Yeast Strain and Maintenancementioning

confidence: 99%

Transcriptomic response of an Antarctic yeast Rhodotorula sp. USM-PSY62 to temperature changes

Abdul Halim, M.,

Chai, C. N.,

Yam, H. C.

et al. 2023

MJM

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Aims: Rhodotorula sp. (USM-PSY62) is a psychrophilic yeast isolated from Antarctic Sea ice that grows optimally at 15 °C. The inevitable global warming poses many challenges to the microbial community in Antarctica. Therefore, this study was conceptualized to observe how USM-PSY62 adapted to fluctuations in temperature. Methodology and results: Rhodotorula sp. (USM-PSY62) was grown in YPD broth until the mid-log phase. Then, the culture was transferred to four different temperatures, specifically at 0 °C, 5 °C, 15 °C and 21 °C for 24 h. Then, the RNA was extracted, sequenced and analysed. During cold adaptation, an elevated transcription of the CorA magnesium transporter gene in USM-PSY62 indicated a higher requirement for magnesium ions to gain additional enzyme cofactors or maintain cytoplasmic fluidity. The HepA homologue coding for DNA/RNA helicase was also over-expressed with log fold change 2.89 in cold conditions possibly to reorganize secondary structures of DNA and RNA. An up-regulation of the catalase gene was also observed, reflecting an increment in the concentration of reactive oxygen species and fluctuations in the associated antioxidant system. The YOP1 gene, which encodes a membrane protein associated with protein transport and membrane traffic, was the most down-regulated, with log2 fold change values of -6.93 lower under cold shock conditions. The genes responsible for the structural maintenance of chromosome (SMC) have a -8.80 in expression log2 fold change, indicating the gene was down-regulated when the temperature was shifted to 0 °C. Upon cold shock, the gene for heat shock factor protein 1 (HSF1) was also down-regulated with a log2 fold change value of -5.86. Hsf1 is a transcriptional regulator which regulates the heat shock responses. Conclusion, significance and impact of study:In conclusion, the transcriptomic responses demonstrated by Rhodotorula sp. USM-PSY62 characterized critical physiological and biochemical compensatory mechanisms especially cellular processes and signalling, information storage and processing, and metabolism to survive at low and high temperatures. This study provides valuable data for industry, especially in the usage of molecular chaperones.

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Constitutive Optimized Production of Streptokinase inSaccharomyces cerevisiaeUtilizing Glyceraldehyde 3-Phosphate Dehydrogenase Promoter ofPichia pastoris

Cited by 15 publications

References 32 publications

Evaluation of clinical efficacy of streptokinase by comparison with the thrombolytic agent on animal model

Evaluation of clinical efficacy of streptokinase by comparison with the thrombolytic agent on animal model

Production of (2R, 3R)-2,3-butanediol using engineered Pichia pastoris: strain construction, characterization and fermentation

Transcriptomic response of an Antarctic yeast Rhodotorula sp. USM-PSY62 to temperature changes

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