A reporter system for prokaryotic and eukaryotic cells based on the thermostable lichenase from Clostridium thermocellum

Пирузян, Е. С.; Goldenkova, I. V.; Musiychuk, Konstantin; Kobets, N. S.; Arman, I.P.; Бобрышева, И. В.; Chekhuta, I. A.; Глазкова, Д. В.

doi:10.1007/s00438-001-0595-8

Cited by 26 publications

(5 citation statements)

References 22 publications

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“…6). The tendency of differences in protein production between promoters was consistent with the results of previous studies (Piruzian et al 2002;Partow et al 2010). However, the display efficiency, i.e., the ratio of yeast cells in the Q2 region, was higher under the GAL1 promoter than the GAP promoter (Fig.…”

Section: Discussionsupporting

confidence: 91%

Evaluation of the yeast surface display system for screening of functional nanobodies

Kajiwara

Aoki

2020

AMB Expr

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Yeast surface display is a powerful technology used to isolate and engineer proteins to improve their activity, specificity, and stability. In this method, gene expression is regulated by promoters, and secretion efficiency is affected by secretion signals. Furthermore, both the accessibility and activity of the displayed proteins are affected by the length of anchor proteins. The ideal promoter, secretion signal, and anchor protein combination depend on the proteins of interest. In this study, we optimized a yeast surface display suitable for nanobody evaluation. We designed five display systems that used different combinations of promoters, secretion signals, and anchor proteins. Anti-hen egg-white lysozyme nanobody was used as the model nanobody. The amount of nanobodies displayed on yeast cells, the number of antigens bound to the displayed nanobodies, and the display efficiency were quantified. Overall, we improved the yeast display system for nanobody engineering and proposed its optimization.

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Section: Discussionsupporting

confidence: 91%

Evaluation of the yeast surface display system for screening of functional nanobodies

Kajiwara

Aoki

2020

AMB Expr

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“…Most of the Lactobacillus and Pediococcus strains in this study were capable of breaking down lichenan and while the lichenase activity was less frequent among the O. oeni, LAB represent a potential source of enzymes for targeting glucans in botrytised wine production. While the genetics and activity of lichenases has been characterised in other bacteria, including Streptococcus bovis (Ekinci et al 1997), Bacillus circulans (Kim 2003), and Clostridium thermocellum (Piruzian et al 2002), to our knowledge, this is the first report of lichenase activity among wine LAB.…”

Section: Discussionmentioning

confidence: 90%

A survey of lactic acid bacteria for enzymes of interest to oenology

Matthews

Grbin

Jiranek

2006

Aust J Grape Wine Res

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Isolates from wine‐associated genera of lactic acid bacteria (LAB), including Lactobacillus, Oenococcus and Pediococcus, were examined for enzymatic properties that could be of value in the production and processing of wine. Up to 438 combinations of bacterial isolates and substrates were examined. Across the 50 isolates investigated, all were found to hydrolyse esters, with the greatest activity being seen amongst the oenococci followed by the lactobacilli and pediococci. Lipase activity was restricted to three Lactobacillus isolates, while all tested isolates displayed tannase activity. Cellulase, β‐glucanase and lichenase activities were observed for almost all Pediococcus and Lactobacillus isolates, while such activities were seen for only a minority of oenococci. No degradation of the xylan substrate was detected. Once confirmed under winelike conditions, our findings will help define potentially interesting genera of LAB for winemaking.

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“…The 1,4‐2,3‐β‐glucanase (lichenase) of the thermophilic bacterium C. thermocellum has been used as a reporter to study the activity of some promoters in plants[19], animals and bacteria[20]. We first proposed to employ this thermophilic enzyme as a reporter for studying the regulation of heat shock promoters of E. coli and cyanobacteria, such as promoters of groES , hspA , and cpn60 genes[17].…”

Section: Discussionmentioning

confidence: 99%

Cyanobacterial leader peptides for protein secretion

Sergeyenko¹,

Los²

2003

FEMS Microbiology Letters

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The leader peptide of the major secreted protein PilA1 of the cyanobacterium Synechocystis sp. strain PCC 6803 and several artificial leader peptides have been used to study secretion of the reporter protein lichenase to the culture medium. The strains of Synechocystis carrying lichenase with the leader sequences of PilA and with the leader sequence of Slr2016 efficiently secreted the reporter protein. The artificial leader sequence that was characterized by the overall positive charge (as PilA1 and Slr2016 leaders) also allowed secretion. The artificial leader with negative charge, however, did not allow secretion of the reporter protein. Moreover, no secreted proteins have been isolated from this strain using conventional techniques for preparation of secreted proteins. These data suggest that the general secretion pathway in cyanobacteria, at least for pilins, recognizes the overall charge of the leader sequences, and operates in a sequence-non-specific manner. ß

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A reporter system for prokaryotic and eukaryotic cells based on the thermostable lichenase from Clostridium thermocellum

Cited by 26 publications

References 22 publications

Evaluation of the yeast surface display system for screening of functional nanobodies

Evaluation of the yeast surface display system for screening of functional nanobodies

A survey of lactic acid bacteria for enzymes of interest to oenology

Cyanobacterial leader peptides for protein secretion

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