Simulation-guided enzyme discovery: A new microbial source of cellobiose 2-epimerase

Xiao, Yaqin; Chen, Qiuming; Shakhnovich, Eugene I.; Zhang, Wenli; Mu, Wanmeng

doi:10.1016/j.ijbiomac.2019.08.075

Cited by 24 publications

(13 citation statements)

References 28 publications

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“…The k cat values of Roin-CE toward cellobiose and lactose were 98.5 ± 2.9 and 117.0 ± 7.7/s, respectively. The K m values of Roin-CE toward cellobiose and lactose were 131.0 ± 11.6 mM and 429.9 ± 57.3 mM, respectively, which were both lower than the corresponding values of Dith-CE, investigated by us previously (Xiao et al, 2019). Thus, Roin-CE had a higher affinity for cellobi-ose and epimerase then did the more stable Dith-CE.…”

Section: Specific Activity and Kinetic Parameters Of Roin-cecontrasting

confidence: 55%

“…The melting temperature of Roin-CE was measured by differential scanning calorimetry to be 57.3°C, which is much lower than for thermostable enzymes. The melting temperatures of Dith-CE (Xiao et al, 2019) and Caob-CE (Chen et al, 2017) 3C). This enzyme is relatively stable and can maintain more than 90% of its initial activity after 1-d production below 40°C.…”

Section: Thermostability Of the Recombinant Enzymementioning

confidence: 99%

“…The activity of Roin-CE at 8°C remained the same during our investigation. Although it is not as stable as enzymes from thermophiles, such as Dith-CE (Xiao et al, 2019) and Casa-CE (Park et al, 2011), Roin-CE is stable enough to work at low temperatures.…”

Section: Thermostability Of the Recombinant Enzymementioning

confidence: 99%

“…Recently Xiao et al (2019) developed a genome mining method combined with molecular dynamics (MD) simulations to discover novel enzymes with desired stabilities. Simulation of MD is a potent tool to study the behavior of the biomacromolecules.…”

Section: Introductionmentioning

confidence: 99%

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Computer-aided search for a cold-active cellobiose 2-epimerase

Chen

Xiao

Zhang

et al. 2020

Journal of Dairy Science

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Cellobiose 2-epimerase (CE) is a promising industrial enzyme that can catalyze bioconversion of lactose to its high-value derivatives, namely epilactose and lactulose. A need exists in the dairy industry to catalyze lactose bioconversions at low temperatures to avoid microbial growth. We focused on the discovery of cold-active CE in this study. A genome mining method based on computational prediction was used to screen the potential genes encoding cold-active enzymes. The CE-encoding gene from Roseburia intestinalis, with a predicted high structural flexibility, was expressed heterologously in Escherichia coli. The catalytic property of the recombinant enzyme was extensively studied. The optimum temperature and pH of the enzyme were 45°C and 7.0, respectively. The specific activity of this enzyme to catalyze conversion of lactose to epilactose was measured to be 77.3 ± 1.6 U/mg. The kinetic parameters, including turnover number (k cat ), Michaelis constant (K m ), and catalytic efficiency (k cat /K m ) using lactose as a substrate were 117.0 ± 7.7 s −1 , 429.9 ± 57.3 mM, and 0.27 mM −1 s −1 , respectively. In situ production of epilactose was carried out at 8°C: 20.9% of 68.4 g/L lactose was converted into epilactose in 4 h using 0.02 mg/mL (1.5 U/mL, measured at 45°C) of recombinant enzyme. The enzyme discovered by this in silico method is suitable for low-temperature applications.

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Section: Specific Activity and Kinetic Parameters Of Roin-cecontrasting

confidence: 55%

Section: Thermostability Of the Recombinant Enzymementioning

confidence: 99%

Section: Thermostability Of the Recombinant Enzymementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Computer-aided search for a cold-active cellobiose 2-epimerase

Chen

Xiao

Zhang

et al. 2020

Journal of Dairy Science

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“…But the discarded lactose without recovery will cause serious environmental pollution (Marwaha and Kennedy 1988). Thus, the further utilization of lactose has become one of the current hot topics (Xiao et al 2019b).…”

Section: Introductionmentioning

confidence: 99%

Sugar alcohols derived from lactose: lactitol, galactitol, and sorbitol

Zhang

Chen

et al. 2020

Appl Microbiol Biotechnol

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Lactose is a common natural disaccharide composed of galactose and glucose molecules. It is mainly found in the whey, the byproduct of cheese and casein industries. As the supply of lactose far exceeds demand, a lot of lactose was discarded as the waste every year, which not only leads to resource waste, but also causes environmental pollution. Therefore, the deep processing of lactose as the feedstock has become a hot research topic. The lactose-derived sugar alcohols, including lactitol, sorbitol, and galactitol, have shown great potential applications not only in food manufacture, but also in pharmaceutical, cosmetic, and material fields. In this paper, we focus on the property, physiological effect, production, and application of the lactose-derived sugar alcohols. Key points• The deep processing of lactose as the feedstock has become a hot research topic.• The lactose-derived sugar alcohols show great application values.• Recent advances in the lactose-derived sugar alcohols are reviewed.

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A novel cellobiose 2‐epimerase from anaerobic halophilic Iocasia fonsfrigidae and its ability to convert lactose in fresh goat milk into epilactose

Eat,

Wulansari,

Ketbot

et al. 2024

J Sci Food Agric

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BACKGROUNDCellobiose 2‐epimerase (CE) has received great attention due to its potential applications in the food and pharmaceutical industries. In this study, a novel CE from mesophilic anaerobic halophilic bacterium Iocasia fonsfrigidae strain SP3‐1 (IfCE) was successfully expressed in Escherichia coli and characterized.RESULTSUnlike other CEs, the purified IfCE shows only epimerization activity toward β‐1,4‐glycosidic linkages of disaccharides, including mannobiose, cellobiose and lactose, but not for monosaccharides, β‐1,4‐glycosidic linkages of trisaccharides and α‐1,4‐glycosidic linkages of disaccharides. Only one epimerization product was obtained from the action of IfCE against mannobiose, cellobiose and lactose. Under optimum conditions, 31.0% of epilactose, a rare and low‐calorie prebiotic sweetener with medicinal and pharmacological properties, was obtained from 10 mg mL−1 lactose. IfCE was highly active against lactose under NaCl concentrations up to 500 mmol L−1, possibly due to the excessive basic (arginine and lysine) and acidic (aspartic and glutamic acids) amino acid residues, which are localized on the surface of the halophilic enzyme structure. These residues may protect the enzyme from Cl− and Na+ ions from the environment, respectively. Under normal conditions, IfCE was able to convert lactose present in fresh goat milk to epilactose with a conversion yield of 31% in 10 min. In addition, IfCE has been investigated as a safe enzyme for human allergen.CONCLUSIONThe results suggested that IfCE is a promising candidate to increase the quality and value of milk and dairy products by converting lactose that causes digestive problems in people with lactose intolerance into epilactose. © 2024 Society of Chemical Industry.

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Simulation-guided enzyme discovery: A new microbial source of cellobiose 2-epimerase

Cited by 24 publications

References 28 publications

Computer-aided search for a cold-active cellobiose 2-epimerase

Computer-aided search for a cold-active cellobiose 2-epimerase

Sugar alcohols derived from lactose: lactitol, galactitol, and sorbitol

A novel cellobiose 2‐epimerase from anaerobic halophilic Iocasia fonsfrigidae and its ability to convert lactose in fresh goat milk into epilactose

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