β-N-Acetylglucosaminidase MthNAG from Myceliophthora thermophila C1, a thermostable enzyme for production of N-acetylglucosamine from chitin

Malgorzata Krolicka, Sandra W.A. Hinz, Martijn J. Koetsier, Gerrit Eggink, Lambertus A.M. van den Broek, Carmen G. Boeriu*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Thermostable enzymes are a promising alternative for chemical catalysts currently used for the production of N-acetylglucosamine (GlcNAc) from chitin. In this study, a novel thermostable β-N-acetylglucosaminidase MthNAG was cloned and purified from the thermophilic fungus Myceliophthora thermophila C1. MthNAG is a protein with a molecular weight of 71 kDa as determined with MALDI-TOF-MS. MthNAG has the highest activity at 50 °C and pH 4.5. The enzyme shows high thermostability above the optimum temperature: at 55 °C (144 h, 75% activity), 60 °C (48 h, 85% activity; half-life 82 h), and 70 °C (24 h, 33% activity; half-life 18 h). MthNAG releases GlcNAc from chitin oligosaccharides (GlcNAc)2–5, p-nitrophenol derivatives of chitin oligosaccharides (GlcNAc)1–3-pNP, and the polymeric substrates swollen chitin and soluble chitosan. The highest activity was detected towards (GlcNAc)2. MthNAG released GlcNAc from the non-reducing end of the substrate. We found that MthNAG and Chitinase Chi1 from M. thermophila C1 synergistically degraded swollen chitin and released GlcNAc in concentration of approximately 130 times higher than when only MthNAG was used. Therefore, chitinase Chi1 and MthNAG have great potential in the industrial production of GlcNAc.

Original languageEnglish
Pages (from-to)7441–7454
JournalApplied Microbiology and Biotechnology
Volume102
Issue number17
Early online date25 Jun 2018
DOIs
Publication statusPublished - Sep 2018

Fingerprint

Acetylglucosaminidase
Chitin
Acetylglucosamine
Enzymes
Chitinases
Oligosaccharides
Half-Life
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Chitosan
Fungi
Molecular Weight
Temperature
Proteins

Keywords

  • Chitin
  • Chitosan
  • Myceliophthora thermophila C1
  • N-Acetylglucosamine
  • β-N-Acetylglucosaminidase

Cite this

@article{7f77049a4c1a40539214f5ac1ea51839,
title = "β-N-Acetylglucosaminidase MthNAG from Myceliophthora thermophila C1, a thermostable enzyme for production of N-acetylglucosamine from chitin",
abstract = "Thermostable enzymes are a promising alternative for chemical catalysts currently used for the production of N-acetylglucosamine (GlcNAc) from chitin. In this study, a novel thermostable β-N-acetylglucosaminidase MthNAG was cloned and purified from the thermophilic fungus Myceliophthora thermophila C1. MthNAG is a protein with a molecular weight of 71 kDa as determined with MALDI-TOF-MS. MthNAG has the highest activity at 50 °C and pH 4.5. The enzyme shows high thermostability above the optimum temperature: at 55 °C (144 h, 75{\%} activity), 60 °C (48 h, 85{\%} activity; half-life 82 h), and 70 °C (24 h, 33{\%} activity; half-life 18 h). MthNAG releases GlcNAc from chitin oligosaccharides (GlcNAc)2–5, p-nitrophenol derivatives of chitin oligosaccharides (GlcNAc)1–3-pNP, and the polymeric substrates swollen chitin and soluble chitosan. The highest activity was detected towards (GlcNAc)2. MthNAG released GlcNAc from the non-reducing end of the substrate. We found that MthNAG and Chitinase Chi1 from M. thermophila C1 synergistically degraded swollen chitin and released GlcNAc in concentration of approximately 130 times higher than when only MthNAG was used. Therefore, chitinase Chi1 and MthNAG have great potential in the industrial production of GlcNAc.",
keywords = "Chitin, Chitosan, Myceliophthora thermophila C1, N-Acetylglucosamine, β-N-Acetylglucosaminidase",
author = "Malgorzata Krolicka and Hinz, {Sandra W.A.} and Koetsier, {Martijn J.} and Gerrit Eggink and {van den Broek}, {Lambertus A.M.} and Boeriu, {Carmen G.}",
year = "2018",
month = "9",
doi = "10.1007/s00253-018-9166-3",
language = "English",
volume = "102",
pages = "7441–7454",
journal = "Applied Microbiology and Biotechnology",
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β-N-Acetylglucosaminidase MthNAG from Myceliophthora thermophila C1, a thermostable enzyme for production of N-acetylglucosamine from chitin. / Krolicka, Malgorzata; Hinz, Sandra W.A.; Koetsier, Martijn J.; Eggink, Gerrit; van den Broek, Lambertus A.M.; Boeriu, Carmen G.

In: Applied Microbiology and Biotechnology, Vol. 102, No. 17, 09.2018, p. 7441–7454.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - β-N-Acetylglucosaminidase MthNAG from Myceliophthora thermophila C1, a thermostable enzyme for production of N-acetylglucosamine from chitin

AU - Krolicka, Malgorzata

AU - Hinz, Sandra W.A.

AU - Koetsier, Martijn J.

AU - Eggink, Gerrit

AU - van den Broek, Lambertus A.M.

AU - Boeriu, Carmen G.

PY - 2018/9

Y1 - 2018/9

N2 - Thermostable enzymes are a promising alternative for chemical catalysts currently used for the production of N-acetylglucosamine (GlcNAc) from chitin. In this study, a novel thermostable β-N-acetylglucosaminidase MthNAG was cloned and purified from the thermophilic fungus Myceliophthora thermophila C1. MthNAG is a protein with a molecular weight of 71 kDa as determined with MALDI-TOF-MS. MthNAG has the highest activity at 50 °C and pH 4.5. The enzyme shows high thermostability above the optimum temperature: at 55 °C (144 h, 75% activity), 60 °C (48 h, 85% activity; half-life 82 h), and 70 °C (24 h, 33% activity; half-life 18 h). MthNAG releases GlcNAc from chitin oligosaccharides (GlcNAc)2–5, p-nitrophenol derivatives of chitin oligosaccharides (GlcNAc)1–3-pNP, and the polymeric substrates swollen chitin and soluble chitosan. The highest activity was detected towards (GlcNAc)2. MthNAG released GlcNAc from the non-reducing end of the substrate. We found that MthNAG and Chitinase Chi1 from M. thermophila C1 synergistically degraded swollen chitin and released GlcNAc in concentration of approximately 130 times higher than when only MthNAG was used. Therefore, chitinase Chi1 and MthNAG have great potential in the industrial production of GlcNAc.

AB - Thermostable enzymes are a promising alternative for chemical catalysts currently used for the production of N-acetylglucosamine (GlcNAc) from chitin. In this study, a novel thermostable β-N-acetylglucosaminidase MthNAG was cloned and purified from the thermophilic fungus Myceliophthora thermophila C1. MthNAG is a protein with a molecular weight of 71 kDa as determined with MALDI-TOF-MS. MthNAG has the highest activity at 50 °C and pH 4.5. The enzyme shows high thermostability above the optimum temperature: at 55 °C (144 h, 75% activity), 60 °C (48 h, 85% activity; half-life 82 h), and 70 °C (24 h, 33% activity; half-life 18 h). MthNAG releases GlcNAc from chitin oligosaccharides (GlcNAc)2–5, p-nitrophenol derivatives of chitin oligosaccharides (GlcNAc)1–3-pNP, and the polymeric substrates swollen chitin and soluble chitosan. The highest activity was detected towards (GlcNAc)2. MthNAG released GlcNAc from the non-reducing end of the substrate. We found that MthNAG and Chitinase Chi1 from M. thermophila C1 synergistically degraded swollen chitin and released GlcNAc in concentration of approximately 130 times higher than when only MthNAG was used. Therefore, chitinase Chi1 and MthNAG have great potential in the industrial production of GlcNAc.

KW - Chitin

KW - Chitosan

KW - Myceliophthora thermophila C1

KW - N-Acetylglucosamine

KW - β-N-Acetylglucosaminidase

U2 - 10.1007/s00253-018-9166-3

DO - 10.1007/s00253-018-9166-3

M3 - Article

VL - 102

SP - 7441

EP - 7454

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

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ER -