Cargando…

AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches

BACKGROUND: Starch is an inexpensive and renewable raw material for numerous industrial applications. However, most starch-based products are not cost-efficient due to high-energy input needed in traditional enzymatic starch conversion processes. Therefore, α-amylase with high efficiency to directly...

Descripción completa

Detalles Bibliográficos
Autores principales: Fang, Wei, Xue, Saisai, Deng, Pengjun, Zhang, Xuecheng, Wang, Xiaotang, Xiao, Yazhong, Fang, Zemin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6477751/
https://www.ncbi.nlm.nih.gov/pubmed/31044008
http://dx.doi.org/10.1186/s13068-019-1432-9
_version_ 1783413073308024832
author Fang, Wei
Xue, Saisai
Deng, Pengjun
Zhang, Xuecheng
Wang, Xiaotang
Xiao, Yazhong
Fang, Zemin
author_facet Fang, Wei
Xue, Saisai
Deng, Pengjun
Zhang, Xuecheng
Wang, Xiaotang
Xiao, Yazhong
Fang, Zemin
author_sort Fang, Wei
collection PubMed
description BACKGROUND: Starch is an inexpensive and renewable raw material for numerous industrial applications. However, most starch-based products are not cost-efficient due to high-energy input needed in traditional enzymatic starch conversion processes. Therefore, α-amylase with high efficiency to directly hydrolyze high concentration raw starches at a relatively lower temperature will have a profound impact on the efficient application of starch. RESULTS: A novel raw starch digesting α-amylase (named AmyZ1) was screened and cloned from a deep-sea bacterium Pontibacillus sp. ZY. Phylogenetic analysis showed that AmyZ1 was a member of subfamily 5 of glycoside hydrolase family 13. When expressed in Escherichia coli, the recombinant AmyZ1 showed high activity at pH 6.0–7.5 and 25–50 °C. Its optimal pH and temperature were 7.0 and 35 °C, respectively. Similar to most α-amylases, AmyZ1 activity was enhanced (2.4-fold) by 1.0 mM Ca(2+). Its half-life time at 35 °C was also extended from about 10 min to 100 min. In comparison, AmyZ1 showed a broad substrate specificity toward raw starches, including those derived from rice, corn, and wheat. The specific activity of AmyZ1 towards raw rice starch was 12,621 ± 196 U/mg, much higher than other reported raw starch hydrolases. When used in raw starch hydrolyzing process, AmyZ1 hydrolyzed 52%, 47% and 38% of 30% (w/v) rice, corn, and wheat starch after 4 h incubation. It can also hydrolyze marine raw starch derived from Chlorella pyrenoidosa, resulting in 50.9 mg/g DW (dry weight of the biomass) of reducing sugars after 4 h incubation at 35 °C. Furthermore, when hydrolyzing raw corn starch using the combination of AmyZ1 and commercial glucoamylase, the hydrolysis rate reached 75% after 4.5 h reaction, notably higher than that obtained in existing starch-processing industries. CONCLUSIONS: As a novel raw starch-digesting α-amylase with high specific activity, AmyZ1 efficiently hydrolyzed raw starches derived from both terrestrial and marine environments at near ambient temperature, suggesting its application potential in starch-based industrial processes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-019-1432-9) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-6477751
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-64777512019-05-01 AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches Fang, Wei Xue, Saisai Deng, Pengjun Zhang, Xuecheng Wang, Xiaotang Xiao, Yazhong Fang, Zemin Biotechnol Biofuels Research BACKGROUND: Starch is an inexpensive and renewable raw material for numerous industrial applications. However, most starch-based products are not cost-efficient due to high-energy input needed in traditional enzymatic starch conversion processes. Therefore, α-amylase with high efficiency to directly hydrolyze high concentration raw starches at a relatively lower temperature will have a profound impact on the efficient application of starch. RESULTS: A novel raw starch digesting α-amylase (named AmyZ1) was screened and cloned from a deep-sea bacterium Pontibacillus sp. ZY. Phylogenetic analysis showed that AmyZ1 was a member of subfamily 5 of glycoside hydrolase family 13. When expressed in Escherichia coli, the recombinant AmyZ1 showed high activity at pH 6.0–7.5 and 25–50 °C. Its optimal pH and temperature were 7.0 and 35 °C, respectively. Similar to most α-amylases, AmyZ1 activity was enhanced (2.4-fold) by 1.0 mM Ca(2+). Its half-life time at 35 °C was also extended from about 10 min to 100 min. In comparison, AmyZ1 showed a broad substrate specificity toward raw starches, including those derived from rice, corn, and wheat. The specific activity of AmyZ1 towards raw rice starch was 12,621 ± 196 U/mg, much higher than other reported raw starch hydrolases. When used in raw starch hydrolyzing process, AmyZ1 hydrolyzed 52%, 47% and 38% of 30% (w/v) rice, corn, and wheat starch after 4 h incubation. It can also hydrolyze marine raw starch derived from Chlorella pyrenoidosa, resulting in 50.9 mg/g DW (dry weight of the biomass) of reducing sugars after 4 h incubation at 35 °C. Furthermore, when hydrolyzing raw corn starch using the combination of AmyZ1 and commercial glucoamylase, the hydrolysis rate reached 75% after 4.5 h reaction, notably higher than that obtained in existing starch-processing industries. CONCLUSIONS: As a novel raw starch-digesting α-amylase with high specific activity, AmyZ1 efficiently hydrolyzed raw starches derived from both terrestrial and marine environments at near ambient temperature, suggesting its application potential in starch-based industrial processes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-019-1432-9) contains supplementary material, which is available to authorized users. BioMed Central 2019-04-23 /pmc/articles/PMC6477751/ /pubmed/31044008 http://dx.doi.org/10.1186/s13068-019-1432-9 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Fang, Wei
Xue, Saisai
Deng, Pengjun
Zhang, Xuecheng
Wang, Xiaotang
Xiao, Yazhong
Fang, Zemin
AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches
title AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches
title_full AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches
title_fullStr AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches
title_full_unstemmed AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches
title_short AmyZ1: a novel α-amylase from marine bacterium Pontibacillus sp. ZY with high activity toward raw starches
title_sort amyz1: a novel α-amylase from marine bacterium pontibacillus sp. zy with high activity toward raw starches
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6477751/
https://www.ncbi.nlm.nih.gov/pubmed/31044008
http://dx.doi.org/10.1186/s13068-019-1432-9
work_keys_str_mv AT fangwei amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches
AT xuesaisai amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches
AT dengpengjun amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches
AT zhangxuecheng amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches
AT wangxiaotang amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches
AT xiaoyazhong amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches
AT fangzemin amyz1anovelaamylasefrommarinebacteriumpontibacillusspzywithhighactivitytowardrawstarches