Purification and characterization of a novel acid phosphatase from the split gill mushroom <i>Schizophyllum commune</i>

Zhang, Guoqing; Chen, Qingjun; Jian, Sun; Wang, Hexiang; Han, Chun-Hua

doi:10.1002/jobm.201200218

Cited by 10 publications

(11 citation statements)

References 36 publications

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“…Our results revealed that the substrate affinities were in the low micromolar range with up to twofold differences; K cat values differed by up to 2.5‐fold. The K M values we determined are lower than those measured for acid phosphatases from different sources using classical spectrophotometric assays (Reilly et al ., 2009; Zhang et al ., 2013; Wang et al ., 2018).…”

Section: Resultsmentioning

confidence: 99%

Developing robust protein analysis profiles to identify bacterial acid phosphatases in genomes and metagenomic libraries

Udaondo

Duque

Daddaoua

et al. 2020

Environmental Microbiology

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Summary Phylogenetic analysis of more than 4000 annotated bacterial acid phosphatases was carried out. Our analysis enabled us to sort these enzymes into the following three types: (1) class B acid phosphatases, which were distantly related to the other types, (2) class C acid phosphatases and (3) generic acid phosphatases (GAP). Although class B phosphatases are found in a limited number of bacterial families, which include known pathogens, class C acid phosphatases and GAP proteins are found in a variety of microbes that inhabit soil, fresh water and marine environments. As part of our analysis, we developed three profiles, named Pfr‐B‐Phos, Pfr‐C‐Phos and Pfr‐GAP, to describe the three groups of acid phosphatases. These sequence‐based profiles were then used to scan genomes and metagenomes to identify a large number of formerly unknown acid phosphatases. A number of proteins in databases annotated as hypothetical proteins were also identified by these profiles as putative acid phosphatases. To validate these in silico results, we cloned genes encoding candidate acid phosphatases from genomic DNA or recovered from metagenomic libraries or genes synthesized in vitro based on protein sequences recovered from metagenomic data. Expression of a number of these genes, followed by enzymatic analysis of the proteins, further confirmed that sequence similarity searches using our profiles could successfully identify previously unknown acid phosphatases.

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Section: Resultsmentioning

confidence: 99%

Developing robust protein analysis profiles to identify bacterial acid phosphatases in genomes and metagenomic libraries

Udaondo

Duque

Daddaoua

et al. 2020

Environmental Microbiology

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“…29–32 More importantly, LDH, GOx and HRP display significant reduction of catalytic activities at 50 °C, while the activity of Lip and AP is rapidly decreases above 40 °C. 8,33–35 Methods to stabilize these enzymes so that they can function at high temperatures are needed to enhance their catalytic activities and is a challenge that should be addressed.…”

Section: Introductionmentioning

confidence: 99%

Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis

et al. 2017

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We report a general and modular approach for the synthesis of multi enzyme–polymer conjugates (MECs) consisting of five different enzymes of diverse isoelectric points and distinct catalytic properties conjugated within a single universal polymer scaffold. The five model enzymes chosen include glucose oxidase (GOx), acid phosphatase (AP), lactate dehydrogenase (LDH), horseradish peroxidase (HRP) and lipase (Lip). Poly(acrylic acid) (PAA) is used as the model synthetic polymer scaffold that will covalently conjugate and stabilize multiple enzymes concurrently. Parallel and sequential synthetic protocols are used to synthesise MECs, 5-P and 5-S, respectively. Also, five different single enzyme–PAA conjugates (SECs) including GOx–PAA, AP–PAA, LDH–PAA, HRP–PAA and Lip–PAA are synthesized. The composition, structure and morphology of MECs and SECs are confirmed by agarose gel electrophoresis, dynamic light scattering, circular dichroism spectroscopy and transmission electron microscopy. The bioreactor comprising MEC functions as a single biocatalyst can carry out at least five different or orthogonal catalytic reactions by virtue of the five stabilized enzymes, which has never been achieved to-date. Using activity assays relevant for each of the enzymes, for example AP, the specific activity of AP at room temperature and 7.4 pH in PB is determined and set at 100%. Interestingly, MECs 5-P and 5-S show specific activities of 1800% and 600%, respectively, compared to 100% specific activity of AP at room temperature (RT). The catalytic efficiencies of 5-P and 5-S are 1.55 × 10−3 and 1.68 × 10−3, respectively, compared to 9.11 × 10−5 for AP under similar RT conditions. Similarly, AP relevant catalytic activities of 5-P and 5-S at 65 °C show 100 and 300%, respectively, relative to native AP activity at RT as the native AP is catalytically inactive at 65 °C The catalytic activity trends suggest: (1) MECs show enhanced catalytic activities compared to native enzymes under similar assay conditions and (2) 5-S is better suited for high temperature biocatalysis, while both 5-S and 5-P are suitable for room temperature biocatalysis. Initial cytotoxicity results show that these MECs are non-lethal to human cells including human embryonic kidney [HEK] cells when treated with doses of 0.01 mg mL−1 for 72 h. This cytotoxicity data is relevant for future biological applications.

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“…Various methods have been used to purify relatively large numbers of protein molecules, while separation is often affected by the differences of the target protein and the properties of other substances present in the sample, such as solubility, precipitation, size, polarity, and the binding affinity of ammonium sulfate/acetone/ethanol precipitation followed by ultrafiltration, ion exchange, and gel filtration chromatography (Vohra and Satyanarayana, 2003;Boyce and Walsh, 2007). Therefore, combinations of two or more methods are commonly used for the purification of fungal phytases (Zhang et al, 2013a; TABLE 1 | Types of fermentation and substrates for fungal phytase production.…”

Section: Purification and Characterization Of Fungal Phytasesmentioning

confidence: 99%

Bioprocess for Production, Characteristics, and Biotechnological Applications of Fungal Phytases

et al. 2020

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Phytases are a group of enzymes that hydrolyze the phospho-monoester bonds of phytates. Phytates are one of the major forms of phosphorus found in plant tissues. Fungi are mainly used for phytase production. The production of fungal phytases has been achieved under three different fermentation methods including solid-state, semi-solid-state, and submerged fermentation. Agricultural residues and other waste materials have been used as substrates for the evaluation of enzyme production in the fermentation process. Nutrients, physical conditions such as pH and temperature, and protease resistance are important factors for increasing phytase production. Fungal phytases are considered monomeric proteins and generally possess a molecular weight of between 14 and 353 kDa. Fungal phytases display a broad substrate specificity with optimal pH and temperature ranges between 1.3 and 8.0 and 37-67 • C, respectively. The crystal structure of phytase has been studied in Aspergillus. Notably, thermostability engineering has been used to improve relevant enzyme properties. Furthermore, fungal phytases are widely used in food and animal feed additives to improve the efficiency of phosphorus intake and reduce the amount of phosphorus in the environment.

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Purification and characterization of a novel acid phosphatase from the split gill mushroom Schizophyllum commune

Cited by 10 publications

References 36 publications

Developing robust protein analysis profiles to identify bacterial acid phosphatases in genomes and metagenomic libraries

Developing robust protein analysis profiles to identify bacterial acid phosphatases in genomes and metagenomic libraries

Nanoarmoring: strategies for preparation of multi-catalytic enzyme polymer conjugates and enhancement of high temperature biocatalysis

Bioprocess for Production, Characteristics, and Biotechnological Applications of Fungal Phytases

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