Overexpression and Purification of Microbial Pro-Transglutaminase from Streptomyces cinnamoneum and in vitro Processing by Streptomyces albogriseolus Proteases.

TAGUCHI, SEIICHI; Arakawa, Kumiko; YOKOYAMA, KEIICHI; Takehana, Shino; Momose, Haruo

doi:10.1263/jbb.94.478

Cited by 5 publications

(8 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, as in the case of the pro-transglutaminase secreted by S. cinnamoneum (32), it was cleaved by SAM-P45 from S. albogriseolus at the C side of 41-Ser of the pro-domain and converted to an active form. We could detect SAM-P45 activity in the supernatant of C. glutamicum harboring a pro-SAM-P45 expression plasmid.…”

Section: Discussionmentioning

confidence: 95%

“…(4) and converts it to an active form (32). Since the amino acid sequences of the pro-transglutaminase from S. cinnamoneum and S. mobaraense are 77% homologous, we tested whether the pro-MTG released by C. glutamicum could be processed by SAM-P45.…”

Section: Sequence Of the S Mobaraense Prepro-mtgmentioning

confidence: 99%

“…Moreover, the prodomain is processed by a subtilisin-like protease from Streptomyces albogriseolus (31,32), when the protease is cosecreted by C. glutamicum, and is converted into active-form MTG. We thus demonstrate that C. glutamicum can efficiently secrete two proteins derived from actinomycetes and that it has potential as a host for industrial-scale protein production.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Secretion of Active-Form Streptoverticillium mobaraense Transglutaminase by Corynebacterium glutamicum : Processing of the Pro-Transglutaminase by a Cosecreted Subtilisin-Like Protease from Streptomyces albogriseolus

Kikuchi

Date

Yokoyama

et al. 2003

Appl Environ Microbiol

124

View full text Add to dashboard Cite

The transglutaminase secreted by Streptoverticillium mobaraense is a useful enzyme in the food industry. A fragment of transglutaminase was secreted by Corynebacterium glutamicum when it was coupled on a plasmid to the promoter and signal peptide of a cell surface protein from C. glutamicum. We analyzed the signal peptide and the pro-domain of the transglutaminase gene and found that the signal peptide consists of 31 amino acid residues and the pro-domain consists of 45 residues. When the pro-domain of the transglutaminase was used, the pro-transglutaminase was secreted efficiently by C. glutamicum but had no enzymatic activity. However, when the plasmid carrying the S. mobaraense transglutaminase also encoded SAM-P45, a subtilisin-like serine protease derived from Streptomyces albogriseolus, the peptide bond to the C side of 41-Ser of the pro-transglutaminase was hydrolyzed, and the pro-transglutaminase was converted to an active form. Our findings suggest that C. glutamicum has potential as a host for industrial-scale protein production.Transglutaminases (protein-glutamine ␥-glutamyltransferase, EC 2.3.2.13) are a family of enzymes that catalyze an acyl transfer reaction between a ␥-carboxyamide group of a glutamine residue in a peptide chain and a ␥-amino group of a lysine residue, resulting in the formation of an ε-(␥-glutamyl) lysine cross-link (6). Transglutaminases are widely distributed, and the physiological properties of several of them have been studied. Transglutaminases derived from animals, for example, human blood coagulation factor XIII, human epidermis keratinocyte transglutaminase, guinea pig liver transglutaminase, and fish liver transglutaminase, are calcium-dependent enzymes (6, 24, 38). Calcium-independent transglutaminases have been discovered in bacteria belonging to the actinomycetes, which include, for example, Streptoverticillium cinnamoneum (4) and Streptoverticillium mobaraense. The enzyme from S. mobaraense has been especially well characterized (1, 36).S. mobaraense transglutaminase (MTG [mature-form transglutaminase]) has been used in the food industry for the modification of proteins (9,13,22). It is used in binding meat or fish and gelled food products such as jelly, yogurt, and cheese. Moreover, it has great potential for use in manufacturing materials found in cosmetics, thermostable microcapsules, and carriers for immobilized enzymes. To date, it is produced by conventional fermentation, but it would be desirable to develop a more efficient system, and a number of reports have described the expression and production of MTG in hostvector systems such as Streptomyces lividans (36) and Escherichia coli (33,39). MTG was secreted in microorganisms such as S. lividans (no more than 0.1 mg/liter) (36) and E. coli (about 5 mg/liter) (33); moreover, it was produced by an inclusion body within E. coli (39). The levels of expression in these studies were low, and it would be very difficult to produce MTG on an industrial scale via an inclusion body.

show abstract

Section: Discussionmentioning

confidence: 95%

Section: Sequence Of the S Mobaraense Prepro-mtgmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Secretion of Active-Form Streptoverticillium mobaraense Transglutaminase by Corynebacterium glutamicum : Processing of the Pro-Transglutaminase by a Cosecreted Subtilisin-Like Protease from Streptomyces albogriseolus

Kikuchi

Date

Yokoyama

et al. 2003

Appl Environ Microbiol

124

View full text Add to dashboard Cite

show abstract

“…The pro-region of TGase from Streptomyces was considered to be essential for correct folding et al 1998) and therefore expressing TGases from Streptomyces in their pro-protein form was given more attention. The pro-transglutaminase from Streptomyces cinnamoneum could be produced in large quantities in Streptomyces lividans, but the pro-protein required activation by adding protease after purification (Taguchi et al 2002). The TGase from Streptoverticillium mobaraense could be secreted by Corynebacterium glutamicum as pro-protein and required activation by a co-expressing protease (Kikuchi et al 2003;Date et al 2004).…”

Section: Transcription Analysis Of Tgase Gene In the Original And Recmentioning

confidence: 99%

“…When TGase was expressed in its mature form, it was difficult to obtain the TGase in its active form in E. coli (Yokoyama et al 2000) or the expression level was low in Streptomyces lividans (Washizu et al 1994). When TGase was expressed in its proprotein form, it needed complicated processes to be activated (Taguchi et al 2002;Kikuchi et al 2003;Date et al 2004;Yurimoto et al 2004). However, it might be convenient to express TGase from Streptomyces in its strain owing to the presence of its own protease, which can convert the pro-protein into the active enzyme (Zotzel et al 2003a,b).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2006

View full text Add to dashboard Cite

The transglutaminase (TGase) gene of Streptomyces fradiae was cloned. It had an ORF of 1242 bp, encoding a presumed prepro-region of 82 amino acids and a mature TGase of 331 amino acids. Enhanced expression of the TGase was achieved by introducing another copy of TGase gene into the original host genome which was driven by the strong constitutive promoter, "ermE up", and shown to be expressed at the mRNA and protein levels. TGase activity in the recombinant strain (3.2 U/ml) was improved 1.3-fold when compared to that normally expressed in the original strain (2.4 U/ml). The specific enzyme activity in the recombinant strain (3.8 U/mg) was double that of the original strain (1.9 U/mg).

show abstract

Surfactant Protein of the Streptomyces Subtilisin Inhibitor Family Inhibits Transglutaminase Activation in Streptomyces hygroscopicus

Zhang

Wang

et al. 2008

J. Agric. Food Chem.

View full text Add to dashboard Cite

Transglutaminase (TGase) is widely used in the food industry for improving protein properties by catalyzing the cross-linking of proteins. In Streptomyces, TGase is secreted as a zymogen, and an activation process has been observed in liquid culture. However, the activation mechanism remains unclear. In the present study, the TGase activation process in Streptomyces hygroscopicus was investigated by biochemical approaches. In a liquid culture, Pro-TGase was secreted and gradually was converted into active TGase during the growth period; however, in a cell-free system in which cells were removed from the liquid culture, TGase activation stalled unexpectedly. Subsequently, the TGase activation process was found to be inhibited by a TGase-activating protease inhibitor (TAPI). N-Terminal amino acid sequencing and a homology search of the purified TAPI revealed that it is a member of the Streptomyces subtilisin inhibitor (SSI) family. Furthermore, it was found that TAPI (0.1 mg/mL) decreased the surface tension of water from 72 to 60 mJ/m2 within 5 min, suggesting that it possesses surface activity. This is the first report that an SSI member functions as a surfactant protein. On the basis of these findings, a model for TAPI-regulated TGase activation process was proposed. This study provides novel insights into the TGase activation process in Streptomyces.

show abstract

Overexpression and Purification of Microbial Pro-Transglutaminase from Streptomyces cinnamoneum and in vitro Processing by Streptomyces albogriseolus Proteases.

Cited by 5 publications

References 16 publications

Secretion of Active-Form Streptoverticillium mobaraense Transglutaminase by Corynebacterium glutamicum : Processing of the Pro-Transglutaminase by a Cosecreted Subtilisin-Like Protease from Streptomyces albogriseolus

Secretion of Active-Form Streptoverticillium mobaraense Transglutaminase by Corynebacterium glutamicum : Processing of the Pro-Transglutaminase by a Cosecreted Subtilisin-Like Protease from Streptomyces albogriseolus

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

Surfactant Protein of the Streptomyces Subtilisin Inhibitor Family Inhibits Transglutaminase Activation in Streptomyces hygroscopicus

Contact Info

Product

Resources

About