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Cloning of a novel thermostable glucoamylase from thermophilic fungus Rhizomucor pusillus and high-level co-expression with α-amylase in Pichia pastoris

Overview of attention for article published in BMC Biotechnology, December 2014
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  • Good Attention Score compared to outputs of the same age (69th percentile)
  • Average Attention Score compared to outputs of the same age and source

Mentioned by

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1 tweeter
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1 patent

Citations

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16 Dimensions

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35 Mendeley
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Title
Cloning of a novel thermostable glucoamylase from thermophilic fungus Rhizomucor pusillus and high-level co-expression with α-amylase in Pichia pastoris
Published in
BMC Biotechnology, December 2014
DOI 10.1186/s12896-014-0114-8
Pubmed ID
Authors

Zhenggui He, Lujia Zhang, Youzhi Mao, Jingchao Gu, Qi Pan, Sixing Zhou, Bei Gao, Dongzhi Wei

Abstract

BackgroundFungal amylase, mainly constitute of fungal ¿-amylase and fungal glucoamylase, are utilized in a broad range of industries, such as starch hydrolysis, food and brewing. Although various ¿-amylase and glucoamylase have been found in various fungi, the amylases from Aspergillus dominate the commercial application. One of the main problems exist with regard to these commercial use of amylases is the relatively low thermal and acid stability. In order to maximize the efficiency of starch processing operations, the operating range of glucoamylase and ¿-amylase, in terms of pH, temperature and compatibility with other enzymes should be improved. So developing fungal amylases with increased thermostability and acid stability which could be used in commercial starch process has been attracting researchers¿ interest continually. Besides, synergetic action of glucoamylase and ¿-amylase could facilitate the degradation of starch. And co-expressing glucoamylase with ¿-amylase in one host could avoid the need to ferment repeatedly and improves cost-effectiveness of the process.ResultsA novel fungal glucoamylase (RpGla) gene encoding a putative protein of 512 amino acid residues was cloned from Rhizomucor pusillus. BLAST analysis revealed that RpGla shared the highest identity of 51% with the Rhizopus oryzae glucoamylase ( ABB77799.1 ). The fungal glucoamylase RpGla was expressed in Pichia pastoris (resulting in KM71/9KGla) with maximum activity of 1237 U ml¿1. The optimum pH and temperature of RpGla were pH 4.0 and 70°C, respectively. Fungal ¿-amylase (RpAmy) gene was also cloned from R. pusillus and transformed into KM71/9KGla, resulted in recombinant yeast KM71/9KGla-Z¿Amy, which harboring the RpGla and RpAmy genes simultaneously. The maximum saccharogenic activity of KM71/9KGla-Z¿Amy was 2218 U ml¿1, which improved 79% compared to KM71/9KGla. Soluble starch hydrolyzed by purified RpGla achieved 43% glucose and 34% maltose. Higher productivity was achieved with a final yield of 48% glucose and 47% maltose catalyzed by purified enzyme preparation produced by KM71/9KGla-Z¿Amy.ConclusionsA novel fungal glucoamylase and fungal ¿-amylase genes were cloned from Rhizomucor pusillus. The two enzymes showed good thermostability and acid stability, and the similar biochemical properties facilitated synergetic action of the two enzymes. A dramatic improvement was seen in amylase activity through co-expressing RpGla with RpAmy in Pichia pastoris. This is the first report of improving activity through co-expression glucoamylase with ¿-amylase in P. pastoris. Besides, fungal glucoamylase and ¿-amylase from R. pusillus were shown as promising candidates for further application in starch hydrolysis.

Twitter Demographics

The data shown below were collected from the profile of 1 tweeter who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 35 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Chile 1 3%
Sweden 1 3%
Unknown 33 94%

Demographic breakdown

Readers by professional status Count As %
Student > Master 10 29%
Student > Ph. D. Student 7 20%
Student > Postgraduate 3 9%
Student > Bachelor 3 9%
Student > Doctoral Student 3 9%
Other 4 11%
Unknown 5 14%
Readers by discipline Count As %
Agricultural and Biological Sciences 14 40%
Biochemistry, Genetics and Molecular Biology 10 29%
Business, Management and Accounting 2 6%
Engineering 2 6%
Chemistry 1 3%
Other 1 3%
Unknown 5 14%

Attention Score in Context

This research output has an Altmetric Attention Score of 4. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 11 February 2021.
All research outputs
#5,417,778
of 17,353,889 outputs
Outputs from BMC Biotechnology
#343
of 857 outputs
Outputs of similar age
#90,380
of 310,799 outputs
Outputs of similar age from BMC Biotechnology
#37
of 54 outputs
Altmetric has tracked 17,353,889 research outputs across all sources so far. This one has received more attention than most of these and is in the 68th percentile.
So far Altmetric has tracked 857 research outputs from this source. They typically receive a little more attention than average, with a mean Attention Score of 6.5. This one has gotten more attention than average, scoring higher than 59% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 310,799 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 69% of its contemporaries.
We're also able to compare this research output to 54 others from the same source and published within six weeks on either side of this one. This one is in the 31st percentile – i.e., 31% of its contemporaries scored the same or lower than it.