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Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser

UPMC 1166 bacterial strain was isolated from SG1 liquid biofertiliser and has proven to have an ability to produce indole acetic acid (IAA) and fixed atmosphere nitrogen. The objectives of this research were to characterise UPMC 1166 isolate, to determine the growth kinetics, and effect of different...

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Main Authors: Mansor, Musliyana, Tan, Geok Hun, Abu Asan, Nor Umaira, Abdul Rahim, Raha
Format: Article
Language:English
Published: Universiti Putra Malaysia Press 2018
Online Access:http://psasir.upm.edu.my/id/eprint/65333/1/07%20JTAS-1159-2017.pdf
http://psasir.upm.edu.my/id/eprint/65333/
http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JTAS%20Vol.%2041%20(3)%20Aug.%202018/07%20JTAS-1159-2017.pdf
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spelling my.upm.eprints.653332018-10-05T09:55:24Z http://psasir.upm.edu.my/id/eprint/65333/ Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser Mansor, Musliyana Tan, Geok Hun Abu Asan, Nor Umaira Abdul Rahim, Raha UPMC 1166 bacterial strain was isolated from SG1 liquid biofertiliser and has proven to have an ability to produce indole acetic acid (IAA) and fixed atmosphere nitrogen. The objectives of this research were to characterise UPMC 1166 isolate, to determine the growth kinetics, and effect of different protectants for preservation. UPMC 1166 were characterised phenotypically and genotypically. The growth kinetics was determined using viable cell count and optical density methods. The effect of different protectants on the viability of UPMC 1166, subjected to freeze–drying and freezing at −80o C, was studied. UPMC 1166 belonged to Gram-positive bacteria (with the size of 0.49 – 0.52 × 1.56 – 2.34 µm), catalase positive, rod-shaped with the arrangement of single or paired bacilli, endospore forming and creamy white pigmentation colonies. Based on API biochemical test kit confirmed that UPMC 1166 was under the Bacillus genus. From BLAST, UPMC 1166 showed pairwise sequence similarity range of 99.0% and is closely related to Bacillus siamensis, Bacillus amyloliquefaciens, Bacillus vallismortis, Bacillus subtilis, Bacillus mojavensis, and Bacillus atrophaeus. 16S rRNA gene sequence used for phylogenetic tree analysis suggested that UPMC 1166 is Bacillus methylotrophicus. To obtain the maximum viability after preservation, it is important to harvest cells during the late logarithmic phase of growth and to choose a suitable protective agent. UPMC 1166 needs approximately 16 h to reach the end of the logarithmic phase; consisting of a lag phase up to 2 h and the logarithmic growth that lasted up to 14 h before entering the stationary phase. During freeze–drying, the maximum protection for UPMC 1166 was achieved by using 10% skimmed milk with 1% sodium glutamate, and 5% trehalose. Maximum protection of cells during −80o C cryopreservation was achieved with 10% dimethyl sulfoxide (DMSO). A suitable selection of protectant seemed to be important to acquire maximum cells viability for long-term preservation. Resistance potential of bacterial strains toward preservation procedures is useful from a research and commercial point of view. Universiti Putra Malaysia Press 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/65333/1/07%20JTAS-1159-2017.pdf Mansor, Musliyana and Tan, Geok Hun and Abu Asan, Nor Umaira and Abdul Rahim, Raha (2018) Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser. Pertanika Journal of Tropical Agricultural Science, 41 (3). pp. 1003-1020. ISSN 1511-3701; ESSN: 2231-8542 http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JTAS%20Vol.%2041%20(3)%20Aug.%202018/07%20JTAS-1159-2017.pdf
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 UPMC 1166 bacterial strain was isolated from SG1 liquid biofertiliser and has proven to have an ability to produce indole acetic acid (IAA) and fixed atmosphere nitrogen. The objectives of this research were to characterise UPMC 1166 isolate, to determine the growth kinetics, and effect of different protectants for preservation. UPMC 1166 were characterised phenotypically and genotypically. The growth kinetics was determined using viable cell count and optical density methods. The effect of different protectants on the viability of UPMC 1166, subjected to freeze–drying and freezing at −80o C, was studied. UPMC 1166 belonged to Gram-positive bacteria (with the size of 0.49 – 0.52 × 1.56 – 2.34 µm), catalase positive, rod-shaped with the arrangement of single or paired bacilli, endospore forming and creamy white pigmentation colonies. Based on API biochemical test kit confirmed that UPMC 1166 was under the Bacillus genus. From BLAST, UPMC 1166 showed pairwise sequence similarity range of 99.0% and is closely related to Bacillus siamensis, Bacillus amyloliquefaciens, Bacillus vallismortis, Bacillus subtilis, Bacillus mojavensis, and Bacillus atrophaeus. 16S rRNA gene sequence used for phylogenetic tree analysis suggested that UPMC 1166 is Bacillus methylotrophicus. To obtain the maximum viability after preservation, it is important to harvest cells during the late logarithmic phase of growth and to choose a suitable protective agent. UPMC 1166 needs approximately 16 h to reach the end of the logarithmic phase; consisting of a lag phase up to 2 h and the logarithmic growth that lasted up to 14 h before entering the stationary phase. During freeze–drying, the maximum protection for UPMC 1166 was achieved by using 10% skimmed milk with 1% sodium glutamate, and 5% trehalose. Maximum protection of cells during −80o C cryopreservation was achieved with 10% dimethyl sulfoxide (DMSO). A suitable selection of protectant seemed to be important to acquire maximum cells viability for long-term preservation. Resistance potential of bacterial strains toward preservation procedures is useful from a research and commercial point of view.
format Article
author Mansor, Musliyana
Tan, Geok Hun
Abu Asan, Nor Umaira
Abdul Rahim, Raha
spellingShingle Mansor, Musliyana
Tan, Geok Hun
Abu Asan, Nor Umaira
Abdul Rahim, Raha
Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser
author_facet Mansor, Musliyana
Tan, Geok Hun
Abu Asan, Nor Umaira
Abdul Rahim, Raha
author_sort Mansor, Musliyana
title Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser
title_short Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser
title_full Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser
title_fullStr Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser
title_full_unstemmed Characterisation and effect of protectants on preservation of Bacillus methylotrophicus UPMC 1166 isolated from liquid biofertiliser
title_sort characterisation and effect of protectants on preservation of bacillus methylotrophicus upmc 1166 isolated from liquid biofertiliser
publisher Universiti Putra Malaysia Press
publishDate 2018
url http://psasir.upm.edu.my/id/eprint/65333/1/07%20JTAS-1159-2017.pdf
http://psasir.upm.edu.my/id/eprint/65333/
http://www.pertanika.upm.edu.my/Pertanika%20PAPERS/JTAS%20Vol.%2041%20(3)%20Aug.%202018/07%20JTAS-1159-2017.pdf
_version_ 1643838284133040128
score 13.211869

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