Cloning of a rumen fungal xylanase gene and purification of the recombinant enzyme via artificial oil bodies

Applied Microbiology and Biotechnology
Je-Ruei LiuCheng-Kang Pai

Abstract

A gene encoding a xylanase, named xynS20, was cloned from the ruminal fungus Neocallimastix patriciarum. The DNA sequence of xynS20 revealed that the gene was 1,008 bp in size and encoded amino acid sequences with a predicted molecular weight of 36 kDa. The amino acid sequence alignment showed that the highest sequence identity (28.4%) is with insect gut xylanase XYL6805. According to the sequence-based classification, a putative conserved domain of glycosyl hydrolase family 11 was detected at the N-terminus of XynS20 and a putative conserved domain of family 1 carbohydrate-binding module (CBM) was observed at the C-terminus of XynS20. An Asn-rich linker sequence was found between the N-terminal catalytic domain and the C-terminal CBM of XynS20. To examine the activity of the gene product, xynS20 gene was cloned as an oleosin-fused protein, expressed in Escherichia coli, affinity-purified by formation of artificial oil bodies, released from oleosin by intein-mediated peptide cleavage, and finally harvested by concentration of the supernatant. The specific activity of purified XynS20 toward oat spelt xylan was 1,982.8 U mg(-1). The recombinant XynS20 was stable in the mild acid pH range from 5.0 to 6.0, and the optimum pH was 6....Continue Reading

References

Dec 1, 1972·The American Journal of Clinical Nutrition·M P Bryant
Aug 1, 1996·Computer Applications in the Biosciences : CABIOS·R D Page
Feb 1, 1997·Applied and Environmental Microbiology·X L LiL G Ljungdahl
Sep 1, 1997·Nucleic Acids Research·S F AltschulD J Lipman
May 7, 1999·Parasite : Journal De La Société Française De Parasitologie·B AkendengueR Hocquemiller
Jul 28, 1999·FEMS Microbiology Reviews·N KulkarniM Rao
Dec 10, 1999·Canadian Journal of Microbiology·J H LiuK J Cheng
Apr 18, 2002·Critical Reviews in Biotechnology·S Subramaniyan, P Prema
Sep 11, 2002·Analytical and Bioanalytical Chemistry·Joachim KönigKurt Vogel
Jan 11, 2003·Proceedings of the National Academy of Sciences of the United States of America·Janne LehtiöTuula T Teeri
May 13, 2004·Journal of Agricultural and Food Chemistry·Chi-Chung PengJason T C Tzen
Jun 9, 2004·Applied and Environmental Microbiology·YaLi BrennanBrian A Steer
Jun 30, 2004·Journal of Molecular Biology·Jannick Dyrløv BendtsenSøren Brunak
Jan 18, 2005·FEMS Microbiology Reviews·Tony CollinsGeorges Feller
Feb 3, 2005·FEMS Microbiology Letters·Ya-Hui HuangRuey-Shyang Hseu
Jun 10, 2005·Applied Microbiology and Biotechnology·M L T M PolizeliD S Amorim
Nov 30, 2006·Cellular and Molecular Life Sciences : CMLS·H Hashimoto

❮ Previous
Next ❯

Citations

Jul 2, 2009·Applied Microbiology and Biotechnology·Sibtain AhmedAmer Jamil
Jun 26, 2009·Applied Microbiology and Biotechnology·Junpei ZhouBin Yao
Aug 20, 2009·Applied Microbiology and Biotechnology·Cheng-Kang PaiJe-Ruei Liu
May 8, 2010·Applied Microbiology and Biotechnology·Skander Elleuche, Stefanie Pöggeler
Jan 27, 2011·Biotechnology Letters·Yifeng Li
Mar 28, 2013·Journal of Chemical Biology·Samuel FurseDavid A Gray
Sep 16, 2010·Genome Research·Lena TasseGabrielle Potocki-Veronese
Nov 10, 2011·Biotechnology Advances·Gabriel PaësJohnny Beaugrand
Feb 11, 2011·Journal of Bioscience and Bioengineering·Chih-Yuan ChuJe-Ruei Liu
Dec 18, 2013·Biotechnology Advances·Ana Sofia PinaAna Cecília A Roque
Apr 22, 2019·Current Opinion in Biotechnology·Casey A HookerKevin V Solomon

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.