Enzymatic and structural characterization of hydrolysis of gibberellin A4 glucosyl ester by a rice β-d-glucosidase

Hua, Yuexuan; Sansenya, Sompong; Saetang, Chiraporn; Wakuta, Shinji; Cairns, James R. Ketudat

doi:10.1016/j.abb.2013.06.005

Cited by 20 publications

(14 citation statements)

References 46 publications

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“…Their kinetic properties, including substrate specificity, were characterized using a recombinant protein expression system (Seshadri et al , ). Among these proteins, Os3BGlu6 efficiently hydrolyzed p ‐nitrophenyl (pNP)‐β‐ d ‐fucoside, pNP‐β‐ d ‐glucoside and pNP‐β‐ d ‐galactoside, but had little activity toward other pNP glycosides, suggesting a preference for short oligosaccharides and hydrophobic glycosides (Hua et al , ). Os3BGlu6 also had the highest hydrolysis activity toward gibberellin A 4 β‐ d ‐glucosyl ester (GA 4 ‐GE), among five recombinantly expressed proteins in the rice glycoside hydrolase family (Hua et al , ).…”

Section: Introductionmentioning

confidence: 99%

“…Among these proteins, Os3BGlu6 efficiently hydrolyzed p ‐nitrophenyl (pNP)‐β‐ d ‐fucoside, pNP‐β‐ d ‐glucoside and pNP‐β‐ d ‐galactoside, but had little activity toward other pNP glycosides, suggesting a preference for short oligosaccharides and hydrophobic glycosides (Hua et al , ). Os3BGlu6 also had the highest hydrolysis activity toward gibberellin A 4 β‐ d ‐glucosyl ester (GA 4 ‐GE), among five recombinantly expressed proteins in the rice glycoside hydrolase family (Hua et al , ). The mutations E178Q , E178A , E394D , E394Q and M251N in this enzyme confirmed its role as an acid/base catalyst as well as a catalytic nucleophile in the hydrolysis of both substrates (Hua et al , ).…”

Section: Introductionmentioning

confidence: 99%

“…Os3BGlu6 also had the highest hydrolysis activity toward gibberellin A 4 β‐ d ‐glucosyl ester (GA 4 ‐GE), among five recombinantly expressed proteins in the rice glycoside hydrolase family (Hua et al , ). The mutations E178Q , E178A , E394D , E394Q and M251N in this enzyme confirmed its role as an acid/base catalyst as well as a catalytic nucleophile in the hydrolysis of both substrates (Hua et al , ). However, little is known about the specific physiological function and the mode of action of Os3BGlu6 in vivo .…”

Section: Introductionmentioning

confidence: 99%

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Chloroplastic Os3BGlu6 contributes significantly to cellular ABA pools and impacts drought tolerance and photosynthesis in rice

Chen

Dong

et al. 2020

New Phytologist

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Cellular abscisic acid (ABA) concentration is determined by both de novo biosynthesis and recycling via b-glucosidase(s). However, which rice b-glucosidase(s) are involved in this process remains unknown. Here, we report on a chloroplastic b-glucosidase isoenzyme, Os3BGlu6, that functions in ABA recycling in rice.Disruption of Os3BGlu6 in rice resulted in dwarfism, lower ABA content in leaves, droughtsensitivity, lower photosynthesis rate and higher intercellular CO 2 concentration. Os3BGlu6 could hydrolyze ABA-GE to ABA in vitro. The reversion and overexpression rice lines restored or increased the drought tolerance as shown by the higher b-glucosidase activity, ABA concentrations and expressions of ABA-and drought-responsive genes. Drought induced Os3BGlu6 to form dimers, and the degree of polymerization correlated well with the increase in cellular ABA concentrations and drought tolerance in rice.Os3BGlu6 was responsive to drought and ABA treatments, and the protein was localized to the chloroplast. Disruption of Os3BGlu6 resulted in the increased stomatal density and impaired stomatal movement. Transcriptomics revealed that disruption of Os3BGlu6 resulted in chloroplastic oxidative stress and lowered Rubisco activity even under normal conditions. Taken together, these results suggest that chloroplastically localized Os3BGlu6 significantly affects cellular ABA pools, thereby affecting drought tolerance and photosynthesis in rice.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Chloroplastic Os3BGlu6 contributes significantly to cellular ABA pools and impacts drought tolerance and photosynthesis in rice

Chen

Dong

et al. 2020

New Phytologist

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“…The At/Os1 cluster representative Os3BGlu6, in contrast to the At/Os4 isoenzymes described above, has little activity on β-1,4-linked oligosaccharides and prefers hydrophobic glycosides [6]. It was recently shown to have relatively high activity toward gibberellin GA4 1-O-acyl glucose ester compared to other rice enzymes [7]. At/Os7 cluster representative Os4BGlu12 was originally identified to act on oligosaccharides, such as those released in wounding, but was recently implicated in the release of the phytohormones tuberonic acid and salicylic acid [8], [9], similar to its close relative Os4BGlu13 (also called rice tuberonic acid β-glucosidase 1, OsTAGG1) [10], [11], suggesting these enzymes could have multiple roles in phytohormone and oligosaccharide metabolism.…”

Section: Introductionmentioning

confidence: 96%

Recombinant Expression and Characterization of the Cytoplasmic Rice β-Glucosidase Os1BGlu4

et al. 2014

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The Os1BGlu4 β-glucosidase is the only glycoside hydrolase family 1 member in rice that is predicted to be localized in the cytoplasm. To characterize the biochemical function of rice Os1BGlu4, the Os1bglu4 cDNA was cloned and used to express a thioredoxin fusion protein in Escherichia coli. After removal of the tag, the purified recombinant Os1BGlu4 (rOs1BGlu4) exhibited an optimum pH of 6.5, which is consistent with Os1BGlu4's cytoplasmic localization. Fluorescence microscopy of maize protoplasts and tobacco leaf cells expressing green fluorescent protein-tagged Os1BGlu4 confirmed the cytoplasmic localization. Purified rOs1BGlu4 can hydrolyze p-nitrophenyl (pNP)-β-d-glucoside (pNPGlc) efficiently (k cat/K m = 17.9 mM−1·s−1), and hydrolyzes pNP-β-d-fucopyranoside with about 50% the efficiency of the pNPGlc. Among natural substrates tested, rOs1BGlu4 efficiently hydrolyzed β-(1,3)-linked oligosaccharides of degree of polymerization (DP) 2–3, and β-(1,4)-linked oligosaccharide of DP 3–4, and hydrolysis of salicin, esculin and p-coumaryl alcohol was also detected. Analysis of the hydrolysis of pNP-β-cellobioside showed that the initial hydrolysis was between the two glucose molecules, and suggested rOs1BGlu4 transglucosylates this substrate. At 10 mM pNPGlc concentration, rOs1BGlu4 can transfer the glucosyl group of pNPGlc to ethanol and pNPGlc. This transglycosylation activity suggests the potential use of Os1BGlu4 for pNP-oligosaccharide and alkyl glycosides synthesis.

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“…In Arabidopsis, the production of active ABA from ABA-GE under abiotic stress conditions were catalyzed by β-glucosidases AtBGLU18 and AtBGLU33 [18,19]. In addition, a few BGLUs in rice have also been demonstrated to hydrolyze salicylic acid glucoside [20], and gibberellic acid glucose ester [21].…”

Section: Introductionmentioning

confidence: 99%

Comprehensive identification and characterization of abiotic stress and hormone responsive glycosyl hydrolase family 1 genes in Medicago truncatula

Yang

Jiang

et al. 2020

Preprint

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Background: β-glucosidases (BGLUs) hydrolyze the β-D-glycosidic bond with retention of anomeric configuration. BGLUs were associated with many aspects of plant physiological processes, in particular biotic and abiotic stresses through the activation of phytohormones and defense compounds. However, no comprehensive and systematic investigation on the stress- or hormone-responsive BGLU proteins had ever been reported in plant.Results: In this study, total 51 BGLU genes of the glycoside hydrolase family 1 were identified in one of the model legume plant Medicago truncatula genome, and they were classified into five distinct clusters. Sequence alignments revealed several conserved and characteristic motifs among these MtBGLU proteins. Analyses of their putative signal peptides and N-glycosylation site suggested that the majority of MtBGLU members have dual targeting to the vacuole/chloroplast. Many regulatory elements possibly related with phytohormones and/or abiotic stresses were identified in MtBGLU genes. Moreover, Microarray and qPCR analyses showed that these MtBGLU genes exhibited distinct expression patterns in various tissues, and in response to different abiotic stress and hormonal treatments. Notably, MtBGLU21, MtBGLU22, MtBGLU28, and MtBGLU30 in cluster I were dramatically activated by NaCl, PEG, IAA, ABA, SA and GA3 treatments.Conclusion: Collectively, our genome-wide characterization, evolutionary analysis, and expression pattern analysis of MtBGLU genes suggested that BGLU proteins play crucial roles in response to various abiotic stresses and hormonal cues in M. truncatula. This systematic analysis provided valuable information for the functional characterization and utilization of these MtBGLU genes in improving stress tolerance in M. truncatula and/or other plant species.

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Enzymatic and structural characterization of hydrolysis of gibberellin A4 glucosyl ester by a rice β-d-glucosidase

Cited by 20 publications

References 46 publications

Chloroplastic Os3BGlu6 contributes significantly to cellular ABA pools and impacts drought tolerance and photosynthesis in rice

Chloroplastic Os3BGlu6 contributes significantly to cellular ABA pools and impacts drought tolerance and photosynthesis in rice

Recombinant Expression and Characterization of the Cytoplasmic Rice β-Glucosidase Os1BGlu4

Comprehensive identification and characterization of abiotic stress and hormone responsive glycosyl hydrolase family 1 genes in Medicago truncatula

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