Cloning of Transglutaminase Gene from Streptomyces fradiae and its Enhanced Expression in the Original Strain

Liu, Xiaoqiu; Yang, Xiuqing; Xie, Fuhong; Qian, Shijun

doi:10.1007/s10529-006-9094-7

Cited by 11 publications

(4 citation statements)

References 14 publications

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“…However, the ermE promoter improved TGase production by 0.8 U/mL [17], and there are no reports for TGase expression with the other strong promoters. It has been found that the endogenous promoter of TGase is recognized in S. lividans , and the yield of the recombinant S. platensis TGase reached 2.22 U/mL [15], which suggests that the endogenous promoter of different TGases or its modified versions could be more efficient for TGase expression by S. lividans in contrast to heterologous strong promoters .…”

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

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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.

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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

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“…It was indicated that the activity (3.2 U/ml) of TGase in recombinant strain rised up to 1.3-fold compared with source strain (2.4 U/ml). However, specific activity of the enzyme (3.8 U/mg) in recombinant strain was twice that of the source organism (1.9 U/mg) (Liu et al, 2006). In another study, the Streptomyces hygroscopicus pro-TGase gene was inserted into the integrative vectors pINA1296 (monocopy) and pINA1297 (multicopy) and transfected to Y. lipolytica Po1g or Po1h strain, respectively.…”

Section: Bacterial Production Of Tgasementioning

confidence: 99%

Bacillus Originated Transglutaminase: Properties and Usage

Gün¹,

Yılmaz²

2022

CTNS

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Enzymes with very important duties, form part of our lives and are useful in various fields. Owing to both the intensity of use and the amount of effective production in standard conditions, the production and use of bacterial-originated enzymes are used continuously in agriculture, health, food and many other industrial areas. Among them, transglutaminases (EC 2.3.2.13) are both intracellular and extracellular enzymes included in the transferase group that catalyse cross-links between proteins. Microbial transglutaminase enzymes are frequently used in the food and pharmaceutical industries to change the functional properties of proteins since it increases viscosity, elasticity and water holding capacity. Particularly the positive effects in meat, dairy, and bakery products such as gelling, increasing mechanical strength, and reducing structural deformations, specifically the texture. It also contributes to reducing the use of additives in diets with low protein and fat content. Furthermore, it reduces the time for cooking processes as well as sensory properties enhancement. Transglutaminase enzymes are also being used in other fields such as tissue culture, biochemical and biomedical research, textile, and leather industries. In this review, a broad perspective is presented on the literature dealing with bacterial transglutaminase studies, especially those belonging to the genus Bacillus. In Bacillus spp., transglutaminase gene was mostly reported in B. subtilis, B. amyloliquefaciens, B. cereus, B. nakamura, B. circulans species and also recently in the whole genome of local Bacillus thuringiensis (Bt) SY49.1 strain (JAHKEZ010000474.1) using the RAST database. It was determined that the Bt transglutaminase gene was 93% identical to that of B. cereus species. The presence of transglutaminase gene in agriculturally indispensable Bt strain can confer them a superficial characteristic due to their water holding capacity in terms of increasing the yield and quality of plant products.

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“…The S. mobaraensis mTGase forms a simple monomer showing overall dimensions of 65 × 59 × 41 Å, made up of ≈ 331 amino acids, with a molecular mass of ≈ 37 kDa and the isoelectric point at pH 8.9 (Ando et al 1989;Kashiwagi et al 2002). The tertiary structure of mTGase has a disk-like structure with a central groove having the active-center with a Cys-Asp-His triad, which is the key to the cross-linking efficiency (Griffin et al 2002;Kashiwagi et al 2002;Liu et al 2006).…”

Section: Structure Of Transglutaminase Of Streptomyces Mobaraensismentioning

confidence: 99%

Transglutaminases: part I—origins, sources, and biotechnological characteristics

Duarte

Matte

Bizarro

et al. 2020

World J Microbiol Biotechnol

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The transglutaminases form a large family of intracellular and extracellular enzymes that catalyze cross-links between protein molecules. Transglutaminases crosslinking properties are widely applied to various industrial processes, to improve the firmness, viscosity, elasticity, and water-holding capacity of products in the food and pharmaceutical industries. However, the extremely high costs of obtaining transglutaminases from animal sources have prompted scientists to search for new sources of these enzymes. Therefore, research has been focused on producing transglutaminases by microorganisms, which may present wider scope of use, based on enzyme-specific characteristics. In this review, we present an overview of the literature addressing the origins, types, reactions, and general characterizations of this important enzyme family. A second review will deal with transglutaminases applications in the area of food industry, medicine, pharmaceuticals and biomaterials, as well as applications in the textile and leather industries.

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Cloning of Transglutaminase Gene from Streptomyces fradiae and its Enhanced Expression in the Original Strain

Cited by 11 publications

References 14 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

Bacillus Originated Transglutaminase: Properties and Usage

Transglutaminases: part I—origins, sources, and biotechnological characteristics

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