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Appl Biochem Biotechnol (2015) 177:700712 DOI 10.1007/s12010-015-1774-0
Dzianis Smirnou1 & Martin Krm1 & Jaromr Kulhnek2 & Martina Hermannov2 & Lenka Bobkov1 & Luk Franke1 & Stanislav Pepeliaev1 & Vladimr Velebn3
Received: 27 April 2015 /Accepted: 21 July 2015 / Published online: 4 August 2015# Springer Science+Business Media New York 2015
Abstract Hyaluronidases (HAases) from yeasts were characterized for the first time. The study elucidated that hyaluronate 4-glycanohydrolase and hyaluronan (HA) lyase can be produced by yeasts. Six yeasts producing HAases were found through express screening of activities. The extracellular HAases from two of the yeast isolates, Pseudozyma aphidis and Cryptococcus laurentii, were characterized among them. P. aphidis HAase hydrolyzed -1,4 glycosidic bonds of HA, yielding even-numbered oligosaccharides with N-acetyl-D-glucosamine at the reducing end. C. laurentii
* Dzianis Smirnou
Martin Krm
Jaromr Kulhnek
Martina Hermannov
Lenka Bobkov
Luk Franke
Stanislav Pepeliaev
Vladimr Velebn
1 Contipro Biotech s.r.o., Doln Dobrou 401, 561 02 Doln Dobrou, Czech Republic
2 Contipro Pharma a.s., Doln Dobrou 401, 561 02 Doln Dobrou, Czech Republic
3 Contipro Group s.r.o., Doln Dobrou 401, 561 02 Doln Dobrou, Czech Republic
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Web End = Characterization of Hyaluronan-Degrading Enzymes from Yeasts
Appl Biochem Biotechnol (2015) 177:700712 701
produced hyaluronan lyase, which cleaved -1,4 glycosidic bonds of HA in -elimination reaction, and the products of HA degradation were different-sized even-numbered oligosaccharides. The shortest detected HA oligomer was dimer. The enzymes pH and temperature optima were pH 3.0 and 3745 C (P. aphidis) and pH 6.0 and 37 C (C. laurentii), respectively. Both HAases showed good thermostability.
Keywords Hyaluronate 4-glycanohydrolase . Lyase . Characteristics . Thermostability. Pseudozyma aphidis . Cryptococcus laurentii
Introduction
Hyaluronidases (HAases) are enzymes that degrade predominantly hyaluronan (HA). These enzymes have been employed therapeutically for many years [1]. Besides, they are useful for the production of pharmaceutically valuable HA oligomers [2, 3].
Until recently, three groups of HAases were reported. The first group is represented by vertebrate and venom HAases (EC 3.2.1.35; glycosyl hydrolase family 56). These non-processive enzymes hydrolyze -1,4 glycosidic bonds of HA yielding tetrasaccharide as...