Isolation and characterization of α ‐amylase encoding gene in Bacillus amyloliquefaciens PAS

Amylolytic bacteria are a source of amylase, which is an essential enzyme to support microalgae growth in the bioreactor for microalgae culture. In a previous study, the highest bacterial isolate to hydrolyze amylum (namely PAS) was successfully isolated from Ranu Pani, Indonesia, and it was identified as Bacillus amyloliquefaciens. That bacterial isolate (B. amyloliquefaciens PAS) also has been proven to accelerate Chlorella vulgaris growth in the mini bioreactor. This study aims to detect, isolate, and characterize the PAS’s α‐amylase encoding gene. This study was conducted with DNA extraction, amplification of α‐amylase gene with polymerase chain reaction (PCR) method with the specific primers, DNA sequencing, phylogenetic tree construction, and protein modeling. The result showed that α‐amylase was successfully detected in PAS bacterial isolate. The α‐amylase DNA fragment was obtained 1,468 bp and that translated sequence has an identity of about 98.3% compared to the B. amylolyquefaciens α‐amylase 3BH4 in the Protein Data Bank (PDB). The predicted 3D protein model of the PAS’s α‐amylase encoding gene has amino acid variations that predicted affect the protein’s structure in the small region. This research will be useful for further research to produce recombinant α‐amylase.

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Isolation and characterization of levansucrase-encoding gene from Bacillus amyloliquefaciens

Gene ◽

10.1016/0378-1119(90)90345-r ◽

1990 ◽

Vol 96 (1) ◽

pp. 89-93 ◽

Cited By ~ 27

Author(s):

Leslie B. Tang ◽

Reijer Lenstra ◽

Torben V. Borchert ◽

Vasantha Nagarajan

Keyword(s):

Bacillus Amyloliquefaciens ◽

Isolation And Characterization ◽

Encoding Gene

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Molecular analysis of the Penicillium marneffei glyceraldehyde-3-phosphate dehydrogenase-encoding gene (gpdA) and differential expression of gpdA and the isocitrate lyase-encoding gene (acuD) upon internalization by murine macrophages

Journal of Medical Microbiology ◽

10.1099/jmm.0.2008/002832-0 ◽

2008 ◽

Vol 57 (11) ◽

pp. 1322-1328 ◽

Cited By ~ 15

Author(s):

Sophit Thirach ◽

Chester R. Cooper ◽

Nongnuch Vanittanakom

Keyword(s):

Differential Expression ◽

Isocitrate Lyase ◽

Glyoxylate Cycle ◽

Evolutionary Relationship ◽

Amino Acid Sequences ◽

Penicillium Marneffei ◽

Macrophage Infection ◽

Dimorphic Fungus ◽

Isolation And Characterization ◽

Encoding Gene

Penicillium marneffei is an intracellular dimorphic fungus that can cause a fatal disseminated disease in human immunodeficiency virus-infected patients. The factors that affect the pathogenicity of this fungus remain unclear. Here, we report the isolation and characterization of the gpdA cDNA and genomic clones encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in P. marneffei. Phylogenetic analysis of GAPDH amino acid sequences demonstrated the evolutionary relationship of P. marneffei to other fungi, including the intracellular pathogen Ajellomyces capsulatus. To assess the central importance of phagocytic cells in defence against P. marneffei infection, we used Northern blotting to investigate the response of the isocitrate lyase-encoding gene (acuD) and gpdA to nutrient deprivation inside macrophages. The results revealed that after macrophage internalization, the gene involved in the glyoxylate cycle, acuD, showed higher expression levels as early as 2 h from the start of co-incubation, and the differential expression could be observed again at 8 h after infection. In contrast, the expression of gpdA was downregulated in the yeast phase, as well as during macrophage infection after 2, 4 and 8 h of infection. The induction of P. marneffei acuD was shown to be coordinated with the downregulation of the glycolytic gpdA gene, implying that the cytoplasmic environment of macrophages is deficient in glucose and the glyoxylate pathway could be used by this pathogen to allow subsistence on two-carbon compounds within the host cell following its intracellular persistence.

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Characterization of a Novel Fructosyltransferase from Lactobacillus reuteri That Synthesizes High-Molecular-Weight Inulin and Inulin Oligosaccharides

Applied and Environmental Microbiology ◽

10.1128/aem.68.9.4390-4398.2002 ◽

2002 ◽

Vol 68 (9) ◽

pp. 4390-4398 ◽

Cited By ~ 104

Author(s):

S. A. F. T. van Hijum ◽

G. H. van Geel-Schutten ◽

H. Rahaoui ◽

M. J. E. C. van der Maarel ◽

L. Dijkhuizen

Keyword(s):

Molecular Weight ◽

High Molecular Weight ◽

Lactobacillus Reuteri ◽

Bacterial Strains ◽

Potential Health ◽

Isolation And Characterization ◽

A Cell ◽

Encoding Gene ◽

First Time

ABSTRACT Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli. When incubated with sucrose, the purified recombinant FTF enzyme produced large amounts of fructo-oligosaccharides (FOS) with β-(2→1)-linked fructosyl units, plus a high-molecular-weight fructan polymer (>107) with β-(2→1) linkages (an inulin). FOS, but not inulin, was found in supernatants of L. reuteri strain 121 cultures grown on medium containing sucrose. Bacterial inulin production has been reported for only Streptococcus mutans strains. FOS production has been reported for a few bacterial strains. This paper reports the first-time isolation and molecular characterization of (i) a Lactobacillus ftf gene, (ii) an inulosucrase associated with a generally regarded as safe bacterium, (iii) an FTF enzyme synthesizing both a high molecular weight inulin and FOS, and (iv) an FTF protein containing a cell wall-anchoring LPXTG motif. The biological relevance and potential health benefits of an inulosucrase associated with an L. reuteri strain remain to be established.

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Isolation and characterization of phosphate solubilizing bacteria from the rhizospheric soil of Ponthenpuzha forest, Pathanamthitta (District), Kerala

Research Journal of Biotechnology ◽

10.25303/168rjbt11021 ◽

2021 ◽

Vol 16 (8) ◽

pp. 110-117

Author(s):

Kannan Abhirami ◽

K. Jayakumar

Keyword(s):

Bacterial Isolate ◽

Phosphate Solubilizing Bacteria ◽

Cymbopogon Citratus ◽

16S Rrna Sequence ◽

Isolation And Characterization ◽

16S Rrna Sequence Analysis ◽

Phosphate Solubilizing ◽

Level Identification

Phosphorous is considered as a major parameter for crop yield. Its availability to plant is independent of its abundance. For the plants to utilize phosphorous, it is to be converted to absorbable form. Here, the part rendered by phosphate solubilizing bacteria is significant for it plays a crucial role in the formation of plant usable phosphate from organic forms. In the present work, an effort had been made to isolate and identify phosphate solubilising bacterial isolate from the rhizhospheric soils of various plants in Ponthenpuzha forest. One of the isolate from Cymbopogon citrates responded positively to Pikovskaya’s medium by producing a halo zone during in vitro culture. Colony features and 16S rRNA sequence analysis identified the isolate as Burkholderia sps. We have reported the presence of genus Burkholderia in the rhizospheric zone of Cymbopogon citratus. Further studies are warranted for species level identification of the isolate.

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