Isolation and some properties of β-galactosidase from the thermophilic bacterium Thermus thermophilus

Maciuńska, Jadwiga; Czyz, Beata; Synowiecki, J.

doi:10.1016/s0308-8146(98)00069-7

Cited by 15 publications

(6 citation statements)

References 12 publications

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“…3 B, maximal pTsbg β-galactosidase activity towards ONPG was found at 85 °C. This optimal temperature is higher than described using the same substrate for other counterparts of the genus Thermus such as T.thermophilus HB8 91 , T.thermophilus HB27 92 , T. aquaticus YT‐1 97 , T. oshimai DSM 12092 96 and is the same reported as optimal to T.thermophilus KNOUC114 β-galactosidase 98 . When compared to other genera of thermophilic bacteria β-galactosidases, pTsbg showed higher optimal temperature than documented for the extremely thermophilic C. saccharolyticus and Marinomonas sp.…”

Section: Resultssupporting

confidence: 47%

Exploring the taxonomical and functional profile of As Burgas hot spring focusing on thermostable β-galactosidases

DeCastro

Doane

Dinsdale

et al. 2021

Sci Rep

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In the present study we investigate the microbial community inhabiting As Burgas geothermal spring, located in Ourense (Galicia, Spain). The approximately 23 Gbp of Illumina sequences generated for each replicate revealed a complex microbial community dominated by Bacteria in which Proteobacteria and Aquificae were the two prevalent phyla. An association between the two most prevalent genera, Thermus and Hydrogenobacter, was suggested by the relationship of their metabolism. The high relative abundance of sequences involved in the Calvin–Benson cycle and the reductive TCA cycle unveils the dominance of an autotrophic population. Important pathways from the nitrogen and sulfur cycle are potentially taking place in As Burgas hot spring. In the assembled reads, two complete ORFs matching GH2 beta-galactosidases were found. To assess their functional characterization, the two ORFs were cloned and overexpressed in E. coli. The pTsbg enzyme had activity towards o-Nitrophenyl-β-d-galactopyranoside (ONPG) and p-Nitrophenyl-β-d-fucopyranoside, with high thermal stability and showing maximal activity at 85 °C and pH 6, nevertheless the enzyme failed to hydrolyze lactose. The other enzyme, Tsbg, was unable to hydrolyze even ONPG or lactose. This finding highlights the challenge of finding novel active enzymes based only on their sequence.

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Section: Resultssupporting

confidence: 47%

Exploring the taxonomical and functional profile of As Burgas hot spring focusing on thermostable β-galactosidases

DeCastro

Doane

Dinsdale

et al. 2021

Sci Rep

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“…Thermus genus was first isolated in the 1960s from thermal springs in Yellowstone National Park (12) and, afterward, in several habitats at environmental conditions of high temperature and pH. Although Thermus aquaticus is the most important species of this genus as a result of its capacity to produce DNA polymerase, other species are producers of proteases (13), aminopeptidase (14), galactosidase (15,16), amylase (1), and kinase (17). The occurrence of esterase and/or lipase activity in bacteria of this genus is also known (18–20), and recently we have ascertained and quantified the capacity of four Thermus strains to produce lipolytic enzymes in submerged cultures (21).…”

Section: Introductionmentioning

confidence: 99%

Production of Thermostable Lipolytic Activity by Thermus Species

Fuciños

Domínguez

Sanromán

et al. 2008

Biotechnol Progress

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A quantitative screening for intra- and extracellular lipolytic activity was performed in submerged cultures of four Thermus strains using two different media (named T or D medium). Major differences in the extracellular lipolytic activity were observed in T medium, the highest values being for Thermus thermophilus HB27 and Thermus aquaticus YT1 strains (18 and 33 U/L, respectively). Two enzymes with lipase/esterase activity were identified in the four Thermus strains by zymogram analysis, with molecular weights of 34 and 62 kDa. No kinetic typification of the enzymes as primary metabolites was possible for any of the Thermus strains, because of the lack of a good fitting of the experimental lipolytic activity production rates to the Luedecking and Piret model. However, a linear relationship was found between the absolute values of biomass and total lipase/esterase activity (sum of intracellular and extracellular). For T. thermophilus HB27, an increase in the aeration rate caused the increase in the production of biomass and, particularly, intracellular lipolytic activity but the extracellular lipolytic activity was not affected except for the series with the strongest oxygen limitation. Transmission electronic microscopy revealed that T. thermophilus HB27 formed rotund bodies surrounded by a common membrane in cultures in the early stationary phase. The results suggest the occurrence of a specific mechanism of lipase/esterase secretion that might be due to the different composition and permeability of the cell membranes and those surrounding the rotund bodies.

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“…These inconveniences can be solved if lactose is hydrolysed to monomeric glucose and galactose by the enzyme b-galactosidase. The sweet syrup from milk whey can be used as a sugar additive and also as a basis for fermentation in food industries (Maciunska et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…Extracellular b-galactosidases from fungi Aspergillus oryzae, Penicillium multicolor, have advantages over the enzyme from other taxonomic groups for industrial applications, in terms of easier isolation from the culture supernatant, higher thermal tolerance and optimum activity at lower pH which can help in avoiding contamination (Ogushi et al, 1980, Takenishi et al, 1983. The intrinsic thermoresistance of fungal b-galactosidases up to 608C is a very important characteristic for technological applications in order to improve the efficiency of milk processing (Maciunska et al, 1998, Cowan et al, 1984.…”

Section: Introductionmentioning

confidence: 99%

An efficient two step purification and molecular characterization of β‐galactosidases from Aspergillus oryzae

Todorova-Balvay¹,

Стоилова²,

Gargova³

et al. 2006

J of Molecular Recognition

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Beta-galactosidases (beta-D-galactoside-galactohydrolases (EC 3.2.1.23), lactases) are important industrial enzymes used for the hydrolysis of lactose from milk and milk whey. These enzymes are produced by different organisms and purified by multi-step procedures. The multi-step purification schemes are cost and time ineffective which can also lead to poor yield, denaturation and loss of enzymatic activity. In our study, extracellular beta-galactosidase from mutant strain Aspergillus oryzaeH26-10-7 was purified by a two step procedure, Metal-ion Affinity Chromatography (IMAC) followed by size-exclusion separation. Purified enzyme was characterized by sodium dodecyl Sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and zymographic analysis. This fungal beta-galactosidase was characterized as a protein corresponding to 113 kDa. Enzyme from mutant strain was found to have five times higher catalytic activity on the synthetic substrate o-nitrophenyl-beta-D-galactopyranoside (ONPG) compared to the wild type enzyme. Moreover, the mutant enzyme was more thermo resistant compared to the wild type. This highly important technological characteristic can be exploited in food industry. Moreover, based on the IMAC patterns of wild type and mutant enzymes, similarities in their His topography were supposed.

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Isolation and some properties of β-galactosidase from the thermophilic bacterium Thermus thermophilus

Cited by 15 publications

References 12 publications

Exploring the taxonomical and functional profile of As Burgas hot spring focusing on thermostable β-galactosidases

Exploring the taxonomical and functional profile of As Burgas hot spring focusing on thermostable β-galactosidases

Production of Thermostable Lipolytic Activity by Thermus Species

An efficient two step purification and molecular characterization of β‐galactosidases from Aspergillus oryzae

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