Purification and Characterization of a<i>β</i>-Glucosidase from<i>Polygonum tinctorium</i>, Which Catalyzes Preferentially the Hydrolysis of Indican

Minami, Yoshiko; Kanafuji, Takeo; Miura, Kazuo

doi:10.1271/bbb.60.147

Cited by 42 publications

(24 citation statements)

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“…We also plan to determine whether BglY's ability to cleave indican could have applications for the production of indigo dye from Polygonum tinctorium. The specific activity of purified BglY on indican is 300 times greater than that reported for an enzyme studied for possible industrial use, Novarom G (24), and the K m of BglY with indican is slightly lower than that reported for the ␤-glucosidase produced natively by P. tinctorium (26). VOL.…”

Section: Discussionmentioning

confidence: 59%

Characterization of an Unusual Cold-Active β-Glucosidase Belonging to Family 3 of the Glycoside Hydrolases from the Psychrophilic Isolate Paenibacillus sp. Strain C7

Shipkowski

Brenchley

2005

Appl Environ Microbiol

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We selected for spore-forming psychrophilic bacteria able to use lactose as a carbon source and one isolate, designated Paenibacillus sp. strain C7, that was phylogenetically related to, but distinct from both Paenibacillus macquariensis and Paenibacillus antarcticus. Some Escherichia coli transformants obtained with genomic DNA from this isolate hydrolyzed X-Gal (5-bromo-4-chloro-3-indoyl-␤-D-galactopyranoside) only below 30°C, an indication of cold-active ␤-galactosidase activity. Sequencing of the cloned insert revealed an open reading frame encoding a 756-amino acid protein that, rather than belonging to a family typically known for ␤-galactosidase activity, belonged to glycoside hydrolase family 3, a family of ␤-glucosidases. Because of this unusual placement, the recombinant enzyme (BglY) was purified and characterized. Consistent with its classification, the enzyme had seven times greater activity with the glucoside substrate ONPGlu (o-nitrophenyl-␤-D-glucopyranoside) than with the galactoside substrate ONPGal (o-nitrophenyl-␤-D-galactopyranoside). In addition, the enzyme had, with ONPGlu, a thermal optimum around 30 to 35°C, activity over a broad pH range (5.5 to 10.9), and an especially low K m (<0.003 mM). Further examination of substrate preference showed that the BglY enzyme also hydrolyzed other aryl-␤-glucosides such as helicin, MUG (4-methylumbelliferyl-␤-D-glucopyranoside), esculin, indoxyl-␤-D-glucoside (a natural indigo precursor), and salicin, but had no activity with glucosidic disaccharides or lactose. These characteristics and substrate preferences make the BglY enzyme unique among the family 3 ␤-glucosidases. The hydrolysis of a variety of aryl-␤-glucosides suggests that the enzyme may allow the organism to use these substrates in the environment and that its low K m on indoxyl-␤-D-glucoside may make it useful for producing indigo.Glycoside hydrolases cleave the bond between two carbohydrates or a carbohydrate moiety and another molecule. The classic Enzyme Classification System (EC) groups glycoside hydrolases by substrate specificity; for instance, the ␤-glucosidases collectively have the designation EC 3.2.1.21. The EC system does not use structural or evolutionary information and is thus less useful for categorizing enzymes when the substrate preferences are not known. Therefore, Henrissat (18) initiated a classification system based on amino acid sequences, hydrophobicity plots, and reaction mechanisms. Under this system, ␤-glucosidases are grouped in two glycoside hydrolase families (GHFs), 1 and 3, and the ␤-galactosidases (EC 3.2.1.23) in four families, 1, 2, 35, and 42. Although this system illustrates enzyme relationships, it does not indicate the natural substrate or function of the enzyme. For example, some GHF 1 ␤-glucosidases possess significant ␤-galactosidase activity (5) and the physiological functions of many of these are unknown.The need for biochemical and physiological studies of microbial glycoside hydrolases is highlighted by the prevalence of open reading frames (OR...

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Section: Discussionmentioning

confidence: 59%

Characterization of an Unusual Cold-Active β-Glucosidase Belonging to Family 3 of the Glycoside Hydrolases from the Psychrophilic Isolate Paenibacillus sp. Strain C7

Shipkowski

Brenchley

2005

Appl Environ Microbiol

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“…Indican has previously been shown to be hydrolyzed by a number of β-glucosidases 41–43 , and because E. coli contains several native β-glucosidases, indican could be susceptible to hydrolysis after production. To measure background hydrolysis, 1 g/L indican was fed to MG1655 E. coli , and the loss of indican was quantified over 4 d (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Employing a biochemical protecting group for a sustainable indigo dyeing strategy

et al. 2018

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Indigo is an ancient dye uniquely capable of producing the signature tones in blue denim; however, the dyeing process requires chemical steps that are environmentally damaging. We describe a sustainable dyeing strategy that not only circumvents the use of toxic reagents for indigo chemical synthesis but also removes the need for a reducing agent for dye solubilization. This strategy utilizes a glucose moiety as a biochemical protecting group to stabilize the reactive indigo precursor indoxyl to form indican, preventing spontaneous oxidation to crystalline indigo during microbial fermentation. Application of a β-glucosidase removes the protecting group from indican, resulting in indigo crystal formation in the cotton fibers. We identified the gene coding for the glucosyltransferase PtUGT1 from the indigo plant Polygonum tinctorium and solved the structure of PtUGT1. Heterologous expression of PtUGT1 in Escherichia coli supported high indican conversion, and biosynthesized indican was used to dye cotton swatches and a garment.

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“…(Gilbert et al, 2004;Maier et al, 1990;Minami et al, 1996;Minami et al, 2000;Minami et al, 1997;Torimoto, 1987). 쪽잎을 인위적으로 찧었을 때 뿐만 아니라 쪽잎이 손상된 경우 에도 손상부위가 공기와 접촉하면 인디칸의 가수분해 가 일어날 수 있고, 분리된 인독실이 산소와 접촉하여 인 디고틴이 발생하게 된다 (Laitonjam & Wangkheirakpam, 2011;Stoker et al, 1998).…”

Section: 이론적 배경unclassified

Analysis of the Pigment Contents of Commercial Indigo Powders and Their Effect on the Color and the Antimicrobial Function of Dyed Cotton Fabrics

Ahn²

2013

Journal of the Korean Society of Clothing and Textiles

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Market available fermented indigo powders of Indian origin (FI1, FI2), Chinese origin (FC1, FC2), and raw indigo powders of Indian origin (R1, R2) were examined using TLC and HPLC analyses to investigate their pigment contents. TLC analysis gave Rf values of 0.81 and 0.72 for blue and red pigments, respectively. All the powder products and the synthetic and natural indigo standards eluted at 6.9 min and 8.3 min in the HPLC chromatograms, and the peaks showed the λmax at 610nm and 542nm, representing indigotin and indirubin, respectively. The pigment content calculated based on the area of indigotin and indirubin peaks in the HPLC chromatograms showed that the indigotin content was higher in FC1 and FC2, while FI1 and FI2 had a higher indirubin content. The relative percentage of indirubin was the highest in R2, but the HPLC peak intensity was quite low. Despite the higher indigotin content in FC1 and FC2, cotton dyed with FI1 and FI2 (versus cotton dyed with FC1 and FC2) showed a higher blue (B) hue, the highest K/S values, and the highest antimicrobial effect.

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Purification and Characterization of aβ-Glucosidase fromPolygonum tinctorium, Which Catalyzes Preferentially the Hydrolysis of Indican

Cited by 42 publications

References 5 publications

Characterization of an Unusual Cold-Active β-Glucosidase Belonging to Family 3 of the Glycoside Hydrolases from the Psychrophilic Isolate Paenibacillus sp. Strain C7

Characterization of an Unusual Cold-Active β-Glucosidase Belonging to Family 3 of the Glycoside Hydrolases from the Psychrophilic Isolate Paenibacillus sp. Strain C7

Employing a biochemical protecting group for a sustainable indigo dyeing strategy

Analysis of the Pigment Contents of Commercial Indigo Powders and Their Effect on the Color and the Antimicrobial Function of Dyed Cotton Fabrics

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