Overproduction of soluble recombinant transglutaminase from Streptomyces netropsis in Escherichia coli

Yu, Yu-Jen; Wu, Shih-Cheng; Chan, Hsun-Chuan; Chen, Yu‐Cheng; Chen, Zong-Yu; Yang, Ming-Te

doi:10.1007/s00253-008-1688-7

Cited by 35 publications

(25 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Because the pro-peptide is essential for the correct folding of TGase, direct expression of mature TGase yields insoluble inclusion bodies [8] or inactive enzyme [9]. Thus, TGase is usually expressed in a pro-TGase form [10, 11]. Due to the absence of activating protease in the host strain, co-expression of heterologous proteases is required to convert pro-TGase into active TGase [12].…”

Section: Introductionmentioning

confidence: 99%

Improving the active expression of transglutaminase in Streptomyces lividans by promoter engineering and codon optimization

Liu

Miao

et al. 2016

BMC Biotechnol

View full text Add to dashboard Cite

BackgroundTransglutaminases (TGase), which are synthesized as a zymogen (pro-TGase) in Streptomyces sp., are important enzymes in the food industry. Because this pro-peptide is essential for the correct folding of Streptomyces TGase, TGase is usually expressed in an inactive pro-TGase form, which is then converted to active TGase by the addition of activating proteases in vitro. In this study, Streptomyces hygroscopicus TGase was actively produced by Streptomyces lividans through promoter engineering and codon optimization.ResultsA gene fragment (tg1, 2.6 kb) that encoded the pro-TGase and its endogenous promoter region, signal peptide and terminator was amplified from S. hygroscopicus WSH03-13 and cloned into plasmid pIJ86, which resulted in pIJ86/tg1. After fermentation for 2 days, S. lividans TK24 that harbored pIJ86/tg1 produced 1.8 U/mL of TGase, and a clear TGase band (38 kDa) was detected in the culture supernatant. These results indicated that the pro-TGase was successfully expressed and correctly processed into active TGase in S. lividans TK24 by using the TGase promoter. Based on deletion analysis, the complete sequence of the TGase promoter is restricted to the region from −693 to −48. We also identified a negative element (−198 to −148) in the TGase promoter, and the deletion of this element increased the TGase production by 81.3 %, in contrast to the method by which S. lividans expresses pIJ86/tg1. Combining the deletion of the negative element of the promoter and optimization of the gene codons, the yield and productivity of TGase reached 5.73 U/mL and 0.14 U/mL/h in the recombinant S. lividans, respectively.ConclusionsWe constructed an active TGase-producing strain that had a high yield and productivity, and the optimized TGase promoter could be a good candidate promoter for the expression of other proteins in Streptomyces.

show abstract

Section: Introductionmentioning

confidence: 99%

Improving the active expression of transglutaminase in Streptomyces lividans by promoter engineering and codon optimization

Liu

Miao

et al. 2016

BMC Biotechnol

View full text Add to dashboard Cite

show abstract

“…Our group have identified and characterized some proteins involved in mechanisms of infection by P. cinnamomi: endo-1,3-beta-glucanase GenBank: AM259651.1 (complete cds) (Meirinho et al 2010), endo-glucanase GenBank: AJ964942.1 (partial cds) (unpublished results), glucanase inhibitor protein (GIP) GenBank: AM259384.1 (complete cds) (Martins et al 2014), necrosis-inducing Phytophthora protein 1 (NPP1) GenBank: AM403130.1 (complete cds) (unpublished results) (Gijzen and Nurnberger 2006;Yu et al 2008), and transglutaminase GenBank: AM403129.1 (unpublished results).…”

Section: Pathogenic Processes In Oomycetesmentioning

confidence: 99%

Scientifically advanced solutions for chestnut ink disease

Choupina

Estevinho

Martins

2014

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

On the north regions of Portugal and Spain, the Castanea sativa Mill. culture is extremely important. The biggest productivity and yield break occurs due to the ink disease, the causal agent being the oomycete Phytophthora cinnamomi. This oomycete is also responsible for the decline of many other plant species in Europe and worldwide. P. cinnamomi and Phytophthora cambivora are considered, by the generality of the authors, as the C. sativa ink disease causal agents. Most Phytophthora species secrete large amounts of elicitins, a group of unique highly conserved proteins that are able to induce hypersensitive response (HR) and enhances plant defense responses in a systemic acquired resistance (SAR) manner against infection by different pathogens. Some other proteins involved in mechanisms of infection by P. cinnamomi were identified by our group: endo-1,3-beta-glucanase (complete cds); exo-glucanase (partial cds) responsible by adhesion, penetration, and colonization of host tissues; glucanase inhibitor protein (GIP) (complete cds) responsible by the suppression of host defense responses; necrosis-inducing Phytophthora protein 1 (NPP1) (partial cds); and transglutaminase (partial cds) which inducts defense responses and disease-like symptoms. In this mini-review, we present some scientifically advanced solutions that can contribute to the resolution of ink disease.

show abstract

“…cinnamoneum (Duran et al, 1998), Streptoverticillium mobaraense (Kikuchi et al, 2003), Streptoverticillium ladakanum (Tellez-Luis et al, 2004), Streptomyces netropsis (Yu et al, 2008), and Streptoverticillium hygroscopicus (Aidaroos et al, 2011). It has also been reported to be found in Bacillus subtilis spores (Liu et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

MICROBIAL TRANSGLUTAMINASE OF INDONESIAN Streptomyces sp. : Screening and Production in several media

Fawzya¹,

Zilda²,

Chaniago

et al. 2016

Squalen Bull. Marine Fisheries Postharvest Biotech.

View full text Add to dashboard Cite

Transglutaminase (TGase), an enzyme that catalyzes the formation of inter-and intra-molecular -(l-glutamyl) lysine (GL) crosslinks, plays an important role in surimi-based products production. The development and the diversification of surimi-based products have recently been getting popular in Indonesia. These surimi-based products can be made from various types of fish. These products generally exhibit good gel strength properties, depending on the fish type and the processing method used. Transglutaminase plays an important role in generating such properties. Fish's endogenous TGase reduces quickly after it is caught and is almost completely destroyed by freezing it, applying exogenous TGase may improve fish's gel forming ability. Microbial transglutaminase (MTGase) can potentially be used to increase gel properties. In this research, a total of 228 Streptomyces strains from marine and terrestrial environments were screened and selected based on their ability to produce MTGase. Strain TTA 02 SDS 14 exhibited the highest activity; and therefore, it was selected for further study. The 16S-rRNA gene analysis showed that it shared 99% similarity to S. thioluteus. In order to optimize MTGase activity, enzyme production was carried out using six different formulas media, designated as media A, B, C, D, E, and F. The result shows that the highest MTGase activity was observed in medium B that contains pepton (1.5%), MgSO 4 .7H 2 0 (0.1%), KH 2 PO 4 (0.5%), Na 2 HPO 4 (0.5%), soybean powder (2%), potato starch (2%), and glucose (1.5%). The MTGase activity reached the highest level (1.45 U/ml) after 4 days of incubation

show abstract

Overproduction of soluble recombinant transglutaminase from Streptomyces netropsis in Escherichia coli

Cited by 35 publications

References 27 publications

Improving the active expression of transglutaminase in Streptomyces lividans by promoter engineering and codon optimization

Improving the active expression of transglutaminase in Streptomyces lividans by promoter engineering and codon optimization

Scientifically advanced solutions for chestnut ink disease

MICROBIAL TRANSGLUTAMINASE OF INDONESIAN Streptomyces sp. : Screening and Production in several media

Contact Info

Product

Resources

About