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Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.

Selected endophytic fungi have been report to be inulin degraders to produce fructose or other oligosaccharides. In this study, the Aspergillus sp. producing inulinase were isolated from selected plant species at Serdang area in Malaysia. Fungal isolates were screened solely based on inulin degradin...

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Main Authors: Shamsuddin, Raba'atun Adawiyah, Mustafa, Shuhaimi, Abd. Manap, Mohd. Yazid, Ali, Abdul Manaf, Noormi, Rosli, Subramaniam, Sreeramanan
Format: Article
Language:English
Published: Springer Netherlands 2011
Online Access:http://psasir.upm.edu.my/id/eprint/22310/
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spelling my.upm.eprints.223102014-10-13T07:52:06Z http://psasir.upm.edu.my/id/eprint/22310/ Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp. Shamsuddin, Raba'atun Adawiyah Mustafa, Shuhaimi Abd. Manap, Mohd. Yazid Ali, Abdul Manaf Noormi, Rosli Subramaniam, Sreeramanan Selected endophytic fungi have been report to be inulin degraders to produce fructose or other oligosaccharides. In this study, the Aspergillus sp. producing inulinase were isolated from selected plant species at Serdang area in Malaysia. Fungal isolates were screened solely based on inulin degrading enzymes production and two isolates named Asf1 and Onf1 were selected as the best inulinase enzyme producers. Genomic DNA of these two isolates were extracted and amplified by polymerase chain reaction (PCR). A 1,341 bp DNA fragment containing inulinase gene was successfully amplified from Asf1 fungal isolate and was named as inu2 gene in this study. Based on the morphological characteristics, rDNA and neighbour-joining phylogenetic analysis, Asf1 fungal isolate could display closely-related to the genus of Aspergillus. The complete sequence designated Asf1 Inu2 gene was successfully obtained via rapid-amplification of cDNA ends-polymerase chain reaction (RACE-PCR). A 2.3 kb DNA fragment encoding endoinulinase, inu2, from Asf1 fungal isolate includes an open reading frame of 1,552 bp with calculated molecular weight of 55,954.1 Da and signal peptide sequence of 23 amino acids. The deduced amino acid sequence of the Asf1 inu2 displayed 97, 96, 69 and 22% identities to that of A. ficuum inu2, A. niger inuB, P. purpurogenum and K. marxianus, respectively. Phylogenetic analysis showed that fungal endo- and exo-inulinases have indepently evolved with the respective hydrolytic activities toward terminal and internal β-(2 → 1)-fructofuranosidic linkages in inulin. Springer Netherlands 2011 Article PeerReviewed Shamsuddin, Raba'atun Adawiyah and Mustafa, Shuhaimi and Abd. Manap, Mohd. Yazid and Ali, Abdul Manaf and Noormi, Rosli and Subramaniam, Sreeramanan (2011) Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp. World Journal of Microbiology and Biotechnology, 27 (9). pp. 2173-2185. ISSN 0959-3993; ESSN: 1573-0972 10.1007/s11274-011-0683-9 English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Selected endophytic fungi have been report to be inulin degraders to produce fructose or other oligosaccharides. In this study, the Aspergillus sp. producing inulinase were isolated from selected plant species at Serdang area in Malaysia. Fungal isolates were screened solely based on inulin degrading enzymes production and two isolates named Asf1 and Onf1 were selected as the best inulinase enzyme producers. Genomic DNA of these two isolates were extracted and amplified by polymerase chain reaction (PCR). A 1,341 bp DNA fragment containing inulinase gene was successfully amplified from Asf1 fungal isolate and was named as inu2 gene in this study. Based on the morphological characteristics, rDNA and neighbour-joining phylogenetic analysis, Asf1 fungal isolate could display closely-related to the genus of Aspergillus. The complete sequence designated Asf1 Inu2 gene was successfully obtained via rapid-amplification of cDNA ends-polymerase chain reaction (RACE-PCR). A 2.3 kb DNA fragment encoding endoinulinase, inu2, from Asf1 fungal isolate includes an open reading frame of 1,552 bp with calculated molecular weight of 55,954.1 Da and signal peptide sequence of 23 amino acids. The deduced amino acid sequence of the Asf1 inu2 displayed 97, 96, 69 and 22% identities to that of A. ficuum inu2, A. niger inuB, P. purpurogenum and K. marxianus, respectively. Phylogenetic analysis showed that fungal endo- and exo-inulinases have indepently evolved with the respective hydrolytic activities toward terminal and internal β-(2 → 1)-fructofuranosidic linkages in inulin.
format Article
author Shamsuddin, Raba'atun Adawiyah
Mustafa, Shuhaimi
Abd. Manap, Mohd. Yazid
Ali, Abdul Manaf
Noormi, Rosli
Subramaniam, Sreeramanan
spellingShingle Shamsuddin, Raba'atun Adawiyah
Mustafa, Shuhaimi
Abd. Manap, Mohd. Yazid
Ali, Abdul Manaf
Noormi, Rosli
Subramaniam, Sreeramanan
Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.
author_facet Shamsuddin, Raba'atun Adawiyah
Mustafa, Shuhaimi
Abd. Manap, Mohd. Yazid
Ali, Abdul Manaf
Noormi, Rosli
Subramaniam, Sreeramanan
author_sort Shamsuddin, Raba'atun Adawiyah
title Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.
title_short Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.
title_full Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.
title_fullStr Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.
title_full_unstemmed Molecular cloning and sequence analysis of an inulinase gene from and Aspergillus sp.
title_sort molecular cloning and sequence analysis of an inulinase gene from and aspergillus sp.
publisher Springer Netherlands
publishDate 2011
url http://psasir.upm.edu.my/id/eprint/22310/
_version_ 1643827790306344960
score 13.211869

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