A novel β-galactosidase from Klebsiella oxytoca ZJUH1705 for efficient production of galacto-oligosaccharides from lactose

Huang, Jin; Zhu, Shengquan; Zhao, Linqi; Chen, Li; Du, Meini; Zhang, Chenglin; Yang, Shang‐Tian

doi:10.1007/s00253-020-10679-9

Cited by 29 publications

(17 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The half-saturation coefficient, K m , is a characteristic constant of the enzyme, which is related to the character of the enzyme rather than to the concentration of enzyme. The K m value approximately represents the affinity of the enzyme and the substrate; that is, the smaller the K m value, the greater the affinity of the enzyme and the substrate (Huang et al, 2020). The K m value of lactose was higher (about 15 times) than that of oNPG under the same assay conditions, suggesting that Gal3149 had a higher affinity for oNPG than for lactose.…”

Section: Substrate Specificity and Kinetic Parametersmentioning

confidence: 95%

Overexpression and characterization of a novel GH4 galactosidase with β-galactosidase activity from Bacillus velezensis SW5

Liu

Wang

et al. 2021

Journal of Dairy Science

View full text Add to dashboard Cite

A novel galactosidase gene (gal3149) was identified from Bacillus velezensis SW5 and heterologously expressed in Escherichia coli BL21 (DE3). The novel galactosidase, Gal3149, encoded by gal3149 in an open reading frame of 1,299 bp, was 433 amino acids in length. Protein sequence analysis showed that Gal3149 belonged to family 4 of glycoside hydrolases (GH4). Gal3149 displayed higher enzyme activity for the substrate 2-nitrophenyl-β-d-galactopyranoside (oNPG) than for 4-nitrophenyl-α-d-galactopyranoside (pNPαG). This is the first time that an enzyme belonging to GH4 has been shown to exhibit β-galactosidase activity. Gal3149 showed optimal activity at pH 8.0 and 50°C, and exhibited excellent thermal stability, with retention of 50% relative activity after incubation at a temperature range of 0 to 50°C for 48 h. Gal3149 activity was significantly improved by K + and Na + , and was strongly or completely inhibited by Ag + , Zn 2+ , Tween-80, Cu 2+ , carboxymethyl cellulose, and oleic acid. The rate of hydrolyzed lactose in 1 mL of milk by 1 U of Gal3149 reached about 50% after incubation for 4 h. These properties lay a solid foundation for Gal3149 in application of the lactose-reduced dairy industry.

show abstract

Section: Substrate Specificity and Kinetic Parametersmentioning

confidence: 95%

Overexpression and characterization of a novel GH4 galactosidase with β-galactosidase activity from Bacillus velezensis SW5

Liu

Wang

et al. 2021

Journal of Dairy Science

View full text Add to dashboard Cite

show abstract

“…and the Brenda‐enzyme database, the K m values of GH42 β‐galactosidases ranged from 0.7 to 13 mM using o NPG as the substrate; therefore, the K m value of BtGal42 was within the range of all characterized GH42 β‐galactosidases. Besides, the catalytic efficiency ( K m /k cat of BtGal42 for o NPG was 242.1 s −1 mM −1 , which was higher than that reported for β‐galactosidases from B. velezensis (101.7 s −1 mM −1 ), 41 K. oxytoca ZJUH1705 (38.8 s −1 mM −1 and 7.4 s −1 mM −1 ), 17 and P. torridus (27.4 s −1 mM −1 ) 39 . The activation energy (Ea) of BtGal42 for the hydrolysis of o NPG was 7.4 kJ/mol, which was lower than that of β‐galactosidases from B. animalis BL‐04 (13.8 kJ/mol) 36 and L. culinaris (8.1 kJ/mol) 45 .…”

Section: Resultsmentioning

confidence: 58%

“…The trimeric conformation was also found in some other GH42 β‐galactosidases, such as those from Anthrobacter sp. 20B, 37 K. oxytoca ZJUH1705, 17 B. bifidum S17, 25 and B. subtilis 6 …”

Section: Resultsmentioning

confidence: 99%

“…For instance, the presence of water in the reaction system may cause the hydrolysis of substrate lactose and the target product. Because of the competition between transgalactosylation and hydrolytic reactions, the catalytic process needs to be early terminated when a 50–70% lactose conversion is reached, resulting in a larger amount of glucose and galactose byproducts, as well as unreacted lactose remaining in a medium 17 . Another competitive reaction in this system is the formation of GOS because the donor of substrate lactose is also an efficient acceptor that can compete with the target substrate as a nucleophile acceptor for the enzyme 18 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A GH42 β‐galactosidase from Bifidobacterium thermophilum: Biochemical characterization and synthesis of galactosyl glycerol by reverse hydrolysis

Duan

Jia

Cao

et al. 2022

Biotech and App Biochem

View full text Add to dashboard Cite

In this study, a β‐galactosidase gene (btgal42) was first cloned from Bifidobacterium thermophilum and successfully expressed in Escherichia coli. The molecular weight of the purified BtGal42 was estimated to be 78 kDa by the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and 225 kDa by the size‐exclusion chromatography, indicating that the native BtGal42 was a homotrimer. BtGal42 belonged to the glycosidase hydrolase family 42, exhibiting the maximum activity at pH of 7 and at temperature of 50°C. The enzyme displayed a strictly specific activity toward substrates with β‐galactosyl linkages at the nonreducing ends, of which the activity on 4‐nitrophenyl‐β‐d‐galactopyranoside was the highest, followed by 2‐nitrophenyl‐β‐d‐galactopyranoside and lactose. Among the tested metals and reagents, BtGal42 showed tolerance in the presence of various organic solvents. Importantly, BtGal42 exhibited a high reverse hydrolysis activity when using galactose as the donor and the di‐alcohol ethylene glycol and the trialcohol glycerol as the acceptors. Under unoptimized reaction conditions, the galactosyl glycerol yield reached 62.2 g/L (galactose conversion rate, 41.2%). This study might provide a feasible method for the biosynthesis of galactosyl glycerol from low‐cost glycerol and galactose, which was associated with high conversion efficiency and few byproducts.

show abstract

“…The obtained results are in compliance with the findings of research conducted on immobilized β-galactosidases from Lactobacillus plantarum HF571129 and Klebsiella oxytoca ZJUH1705 which were exploited for producing galacto-oligosaccharides and lactose hydrolysis. 26,27…”

Section: Kinetic Parameters Measurementsmentioning

confidence: 99%

Validation and Optimization of Polyvinyl Alcohol-Functionalized Gold Nanoparticles for Producing Lactose-Free Dairy Products

Alshanberi

Ansari²

2021

Orient. J. Chem

View full text Add to dashboard Cite

The present study demonstrates the synthesis of lactose-free dairy items by Kluyveromyces lactis β-galactosidase bound to polyvinyl alcohol (PVA)-modified gold nanoparticles (AuNPs). The size of AuNPs was analyzed by dynamic light scattering experiment. The developed AuNPs served as a stable matrix for enzyme immobilization which was observed by obtaining 88% immobilization yield. Km and Vmax were determined for soluble and immobilized enzyme by incubating them with varying concentrations of substrate. Our findings demonstrated that immobilization leads to an increase of Km and a decline in Vmax values for the enzyme attached to PVA-functionalized AuNPs. Moreover, the enzyme conjugated to surface functionalized AuNPs displayed exceptional conversion of lactose hydrolysis in batch reactors at 40 oC in contrast to its hydrolysis at 50 oC. Hence, the developed nanosystem [β-galactosidase-(PVA-modified AuNPs)] serves as an excellent model for suggesting its application in other biomedical applications, particularly for constructing lactose based biosensors.

show abstract

A novel β-galactosidase from Klebsiella oxytoca ZJUH1705 for efficient production of galacto-oligosaccharides from lactose

Cited by 29 publications

References 31 publications

Overexpression and characterization of a novel GH4 galactosidase with β-galactosidase activity from Bacillus velezensis SW5

Overexpression and characterization of a novel GH4 galactosidase with β-galactosidase activity from Bacillus velezensis SW5

A GH42 β‐galactosidase from Bifidobacterium thermophilum: Biochemical characterization and synthesis of galactosyl glycerol by reverse hydrolysis

Validation and Optimization of Polyvinyl Alcohol-Functionalized Gold Nanoparticles for Producing Lactose-Free Dairy Products

Contact Info

Product

Resources

About