Constitutive Expression of Thermobifida fusca Thermostable Acetylxylan Esterase Gene in Pichia pastoris

Yang, Chao‐Hsun; Lin, Kuei‐Ying; Chen, Gen‐Hung; Chen, Yu-Fen; Chen, Cheng-Yu; Chen, Weilin; Huang, Yu‐Chun

doi:10.3390/ijms11125143

Cited by 11 publications

(4 citation statements)

References 27 publications

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“…Acetylxylan esterase cloned from T. fusca NTU22 (TfAXE) also exhibits high similarity to cutinases (Huang et al 2010). T. fusca cutinases are the most researched among actinomycete cutinases, which are expressed in hosts other than Escherichia coli , such as Streptomyces rimosus (Sinsereekul et al 2010), Streptomyces lividans (Li et al 2013), Bacillus megaterium (Yang et al 2007), and Pichia pastoris X-33 (TfAXE; Yang et al 2010).…”

Section: Biochemical Bases For Pet Hydrolasesmentioning

confidence: 99%

Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields

Kawai

Kawabata

Oda

2019

Appl Microbiol Biotechnol

436

377

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Enzymatic hydrolysis of polyethylene terephthalate (PET) has been the subject of extensive previous research that can be grouped into two categories, viz. enzymatic surface modification of polyester fibers and management of PET waste by enzymatic hydrolysis. Different enzymes with rather specific properties are required for these two processes. Enzymatic surface modification is possible with several hydrolases, such as lipases, carboxylesterases, cutinases, and proteases. These enzymes should be designated as PET surface–modifying enzymes and should not degrade the building blocks of PET but should hydrolyze the surface polymer chain so that the intensity of PET is not weakened. Conversely, management of PET waste requires substantial degradation of the building blocks of PET; therefore, only a limited number of cutinases have been recognized as PET hydrolases since the first PET hydrolase was discovered by Müller et al. ( Macromol Rapid Commun 26:1400–1405, 2005 ). Here, we introduce current knowledge on enzymatic degradation of PET with a focus on the key class of enzymes, PET hydrolases, pertaining to the definition of enzymatic requirements for PET hydrolysis, structural analyses of PET hydrolases, and the reaction mechanisms. This review gives a deep insight into the structural basis and dynamics of PET hydrolases based on the recent progress in X-ray crystallography. Based on the knowledge accumulated to date, we discuss the potential for PET hydrolysis applications, such as in designing waste stream management. Electronic supplementary material The online version of this article (10.1007/s00253-019-09717-y) contains supplementary material, which is available to authorized users.

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Section: Biochemical Bases For Pet Hydrolasesmentioning

confidence: 99%

Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields

Kawai

Kawabata

Oda

2019

Appl Microbiol Biotechnol

436

377

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“…However, this application may be viewed as controversial as mutants are more effective than wild-type and should be cloned in appropriate hosts. The expression of PET hydrolase genes in hosts other than conventional Escherichia coli has been documented: T. fusca cutinases in Streptomyces remosus , Streptomyces lividans , B. megaterium , and Pichia pastoris X-33 and LC-cutinase in P. pastoris . Use of the produced recombinant microbes/enzymes must be carefully evaluated and balanced with the safety of the ecosystem.…”

Section: Improvement and Prospective Applications Of Pet Hydrolasesmentioning

confidence: 99%

Current State and Perspectives Related to the Polyethylene Terephthalate Hydrolases Available for Biorecycling

Kawai

Kawabata

Oda

2020

ACS Sustainable Chem. Eng.

211

195

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Polyethylene terephthalate (PET) hydrolase is a challenging target as PET is a commonly used plastic that is extremely resistant to enzymatic attack. Since the discovery of a PET hydrolase from Thermobif ida f usca in 2005, novel PET hydrolases and their availability toward waste PET have been investigated. At present, at least four thermophilic cutinases are known as PET hydrolases that could be used for the management of amorphous PET waste, such as packaging materials. Heat-labile PETase from Ideonella sakaiensis and its homologues from mesophilic bacteria exist in the environment. However, PET can be efficiently hydrolyzed with thermophilic hydrolases. This Review focuses on the current state of PET hydrolases and the potential of their application. Contrary to an amorphous PET, the enzymatic hydrolysis of crystalline PET (particularly PET bottles) remains to be fully elucidated. It cannot be assured whether the biorecycling of general PET would be put into practice in the near future, but the plan is getting closer to the goal. PET hydrolases can be versatile polyesterases as they can hydrolyze not only PET but also other polyesters. Additionally, the thermostability of PET hydrolases is advantageous to their application in terms of reaction speed and durability.

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“…Actinomycetes constitute a formidable group of industrially important microorganisms that have been explored for the production of a variety of enzymes. A large number of enzymes have been found and isolated from actinomycetes, including phospholipase [1], phosphatase [2], cellulase [3], keratinase [4], protease [5,6], α-amylase [7,8], and xylanase [9–11], and some genes of them were cloned and expressed in Escherichia coli [12–16], Pichia pastoris [7,12,17] and Streptomyces lividans [1,13,14,18]. Comparision of the productions of these proteins in the three hosts revealed that the expression of active proteins achieved in P. pastoris and S. lividans was much higher than that achieved in E. coli , even expression of some actinomycetes enzymes in E. coli was unsuccessful.…”

Section: Introductionmentioning

confidence: 99%

High-Level Overproduction of Thermobifida Enzyme in Streptomyces lividans Using a Novel Expression Vector

Zhao

et al. 2013

IJMS

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In this study, we constructed a novel Streptomyces-E.coli shuttle vector pZRJ362 combining the xylose isomerase promoter and amylase terminator. A gene encoding the endoglucanase Cel6A in Thermobifida fusca was amplified by PCR, cloned into Streptomyces lividans host strain using the novel expression vector and Pichia pastoris GS115 host strain using the vector pPICZα-C, respectively. Afterwards, the expression pattern and the maximum expression level were comparatively studied in both expression systems. The maximum enzyme activity of Cel6A-(His)6 secreted in S. lividans supernatant after 84-h of cultivation amounted to 5.56 U/mL, which was dramatically higher than that secreted in P. pastoris about 1.4 U/mL after 96-h of cultivation. The maximum expression level of Cel6A-(His)6 in S. lividans supernatant reached up to 173 mg/L after 84-h of cultivation. The endoglucanase activity staining SDS-PAGE showed that there were some minor proteins in S. lividans supernatant which may be the Cel6A derivant by proteolytic degradation, while there was no proteolytic product detected in supernatant of P. pastoris.

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Constitutive Expression of Thermobifida fusca Thermostable Acetylxylan Esterase Gene in Pichia pastoris

Cited by 11 publications

References 27 publications

Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields

Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields

Current State and Perspectives Related to the Polyethylene Terephthalate Hydrolases Available for Biorecycling

High-Level Overproduction of Thermobifida Enzyme in Streptomyces lividans Using a Novel Expression Vector

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