Characterization of Sphingomonas sp. JB13 exo-inulinase: a novel detergent-, salt-, and protease-tolerant exo-inulinase

Abstract

A glycoside hydrolase family 32 exo-inulinase gene was cloned from Sphingomonas sp. JB13 and expressed in Escherichia coli BL21 (DE3). The purified recombinant enzyme (rInuAJB13) showed an apparently optimal activity at pH 5.5 and 55 °C and remained activity at 10-70 °C. The addition of most metal ions and chemical reagents showed little or no effect (retaining more than 76.5 % activity) on the enzyme activity, notably the addition of surfactants SDS, CTAB, Tween 80, and Triton X-100. Most local liquid detergents, including Balin, Walch, Ariel, Tide, Tupperware, and Bluemoon, also showed little or no effect (retaining more than 77.8 % activity) on the enzyme activity. rInuAJB13 exhibited 135.3-163.6 % activity at the NaCl concentration of 1.0-4.5 M. After incubation with up to 57.0 mg mL(-1) trypsin and 90.0 mg mL(-1) proteinase K at 37 °C for 60 min (pH 7.2), rInuAJB13 retained more than 80 % of its initial activity. The enzyme presents a high proportion (28.0 %) of amino acid residues G, A, and V. This paper is the first to report a detergent-, salt-, and protease-tolerant exo-inulinase.

Expression of exo‐inulinase gene from Aspergillus niger 12 in E. coli strain Rosetta‐gami B (DE3) and its characterization

“…The S/I ratio in this study 3.9‐fold lower than our current study. Other studies as in Table also indicate that the S/I ratios found in our study are within the range of values obtained by other investigators. The recombinant exo‐inulinase in our study displays typical inulinase characteristics since its S/I ratio falls in the range of 0.5 − 18.5, whereas true invertases display S/I ratios of several thousand …”

Continuous generation of fructose from Taraxacum officinale tap root extract and inulin by immobilized inulinase in a packed-bed reactor

“…These results are in accordance with those noticed for exo-inulinases of Aspergillus fumigatus, Aspergillus ficuum and Sphingomonas sp. JB13 (Chen et al, 2009;Gill et al, 2006a;Zhou et al, 2015).…”

Glycoside Hydrolase Family 39 β-Xylosidase of Sphingomonas Showing Salt/Ethanol/Trypsin Tolerance, Low-pH/Low-Temperature Activity, and Transxylosylation Activity

“…To obtain the aforementioned β-xylosidase, one may mine resources from new microorganisms. The Sphingomonas genus includes numerous bacterial strains that can survive in various aqueous and terrestrial habitats, plant root systems, clinical specimens, and other environments due to their versatile biodegradative capabilities. , Our previous studies have shown that Sphingomonas sp. JB13 is a new strain that can produce enzymes with interesting properties appropriate for application in food and beverage production that necessitate salt/protease tolerance. , In this study, GH 39 β-xylosidase, designated JB13GH39, is cloned from Sphingomonas sp.…”

“…To obtain the aforementioned β-xylosidase, one may mine resources from new microorganisms. The Sphingomonas genus includes numerous bacterial strains that can survive in various aqueous and terrestrial habitats, plant root systems, clinical specimens, and other environments due to their versatile biodegradative capabilities 11,12. Our previous studies have shown that Sphingomonas sp.…”

“…Our previous studies have shown that Sphingomonas sp. JB13 is a new strain that can produce enzymes with interesting properties appropriate for application in food and beverage production that necessitate salt/protease tolerance 11,12. In this study, GH 39 β-xylosidase, designated JB13GH39, is cloned from Sphingomonas sp.…”