Construction of a Citrinin Gene Cluster Expression System in Heterologous Aspergillus oryzae

2008 ◽  
Vol 106 (5) ◽  
pp. 466-472 ◽  
Author(s):  
Kanae Sakai ◽  
Hiroshi Kinoshita ◽  
Takeo Shimizu ◽  
Takuya Nihira
Keyword(s):  
Gene Cluster ◽  
Aspergillus Oryzae ◽  
Expression System

scholarly journals Development of a Homologous Expression System for and Systematic Site-Directed Mutagenesis Analysis of Thurincin H, a Bacteriocin Produced by Bacillus thuringiensis SF361

2014 ◽  
Vol 80 (12) ◽  
pp. 3576-3584 ◽  
Author(s):  
Gaoyan Wang ◽  
David C. Manns ◽  
John J. Churey ◽  
Randy W. Worobo
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Bacillus Thuringiensis ◽  
Gene Cluster ◽  
Expression Vector ◽  
Expression System ◽  
Structural Genes ◽  
Cry Protein ◽  
Content Type ◽  
Thurincin H

ABSTRACTThurincin H is an antimicrobial peptide produced byBacillus thuringiensisSF361. With a helical back bone, the 31 amino acids of thurincin H form a hairpin structure maintained by four pairs of very unique sulfur-to-α-carbon thioether bonds. The production of thurincin H depends on a putative gene cluster containing 10 open reading frames. The gene cluster includes three tandem structural genes (thnA1,thnA2, andthnA3) encoding three identical 40-amino-acid thurincin H prepeptides and seven other genes putatively responsible for prepeptide processing, regulation, modification, exportation, and self-immunity. A homologous thurincin H expression system was developed by transforming a thurincin H-deficient host with a novel expression vector, pGW133. The host, designatedB. thuringiensisSF361 ΔthnA1ΔthnA2ΔthnA3, was constructed by deletion of the three tandem structural genes from the chromosome of the natural thurincin H producer. The thurincin H expression vector pGW133 was constructed by cloning the thurincin H native promoter,thnA1, and a Cry protein terminator into theEscherichia coli-B. thuringiensisshuttle vector pHT315. Thirty-three different pGW133 variants, each containing a different point mutation in thethnA1gene, were generated and separately transformed intoB. thuringiensisSF361 ΔthnA1ΔthnA2ΔthnA3. Those site-directed mutants contained either a single radical or conservative amino acid substitution on the thioether linkage-forming positions or a radical substitution on all other nonalanine amino acids. The bacteriocin activities ofB. thuringiensisSF361 ΔthnA1ΔthnA2ΔthnA3carrying different pGW133 variants against three different indicator strains were subsequently compared.

scholarly journals Heterologous Biosynthesis and Genomics-Driven Derivatization of Fungal Bioactive Sesterterpenoid Variecolin

2021 ◽  
Author(s):  
Dexiu Yan ◽  
Jemma Arakelyan ◽  
Teng Wan ◽  
Tsz Ki Chan ◽  
Dohyun Ahn ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Anticancer Activity ◽  
Gene Cluster ◽  
Aspergillus Oryzae ◽  
Biosynthetic Gene Cluster ◽  
Heterologous Production ◽  
Biosynthetic Gene ◽  
Aspergillus Aculeatus ◽  
Heterologous Biosynthesis

The biosynthetic gene cluster of fungal bioactive sesterterpenoids, variecolin (1) and variecolactone (2), was identified in Aspergillus aculeatus ATCC 16872. Heterologous production of 1 and 2 was achieved in Aspergillus oryzae by expressing the sesterterpene synthase VrcA and the cytochrome P450 VrcB. Intriguingly, the replacement of VrcB with homologous P450s from other fungal terpenoid pathways yielded three new variecolin analogues, one of which exhibited potent anticancer activity comparable to that of 1.

Efficient Reconstitution of Basidiomycota Diterpene Erinacine Gene Cluster in Ascomycota Host Aspergillus oryzae Based on Genomic DNA Sequences

2019 ◽  
Vol 141 (39) ◽  
pp. 15519-15523 ◽  
Author(s):  
Chengwei Liu ◽  
Atsushi Minami ◽  
Taro Ozaki ◽  
Jing Wu ◽  
Hirokazu Kawagishi ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Aspergillus Oryzae ◽  
Dna Sequences ◽  
Genomic Dna

Analysis of the ionic interaction between the hydrophobin RodA and two cutinases of Aspergillus nidulans obtained via an Aspergillus oryzae expression system

2016 ◽  
Vol 101 (6) ◽  
pp. 2343-2356 ◽  
Author(s):  
Takumi Tanaka ◽  
Mayumi Nakayama ◽  
Toru Takahashi ◽  
Kei Nanatani ◽  
Youhei Yamagata ◽  
...  
Keyword(s):  
Aspergillus Nidulans ◽  
Aspergillus Oryzae ◽  
Expression System ◽  
Ionic Interaction

Aspergillus oryzae strains with a large deletion of the aflatoxin biosynthetic homologous gene cluster differentiated by chromosomal breakage

2006 ◽  
Vol 72 (2) ◽  
pp. 339-345 ◽  
Author(s):  
Yun-Hae Lee ◽  
Mihoko Tominaga ◽  
Risa Hayashi ◽  
Kazutoshi Sakamoto ◽  
Osamu Yamada ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Aspergillus Oryzae ◽  
Large Deletion ◽  
Homologous Gene ◽  
Chromosomal Breakage

scholarly journals Expression of the Clostridium botulinum A2 Neurotoxin Gene Cluster Proteins and Characterization of the A2 Complex

2009 ◽  
Vol 76 (1) ◽  
pp. 40-47 ◽  
Author(s):  
Guangyun Lin ◽  
William H. Tepp ◽  
Christina L. Pier ◽  
Mark J. Jacobson ◽  
Eric A. Johnson
Keyword(s):  
Gene Cluster ◽  
Clostridium Botulinum ◽  
Polyclonal Antibodies ◽  
Expression System ◽  
Middle Part ◽  
Open Reading Frames ◽  
Protein Sequencing ◽  
Medium Size ◽  
Reading Frames

ABSTRACT Clostridium botulinum subtype A2 possesses a botulinum neurotoxin type A (BoNT/A) gene cluster consisting of an orfX cluster containing open reading frames (ORFs) of unknown functions. To better understand the association between the BoNT/A2 complex proteins, first, the orfX cluster proteins (ORFX1, ORFX3, P47, and the middle part of NTNH) from C. botulinum A2 strain Kyoto F and NTNH of A1 strain ATCC 3502 were expressed by using either an Escherichia coli or a C. botulinum expression system. Polyclonal antibodies against individual orfX cluster proteins were prepared by immunizing a rabbit and mice against the expressed proteins. Antibodies were then utilized as probes to determine which of the A2 orfX cluster genes were expressed in the native A2 culture. N-terminal protein sequencing was also employed to specifically detect ORFX2. Results showed that all of the neurotoxin cluster proteins, except ORFX1, were expressed in the A2 culture. A BoNT/A2 toxin complex (TC) was purified which showed that C. botulinum A2 formed a medium-size (300-kDa) TC composed of BoNT/A2 and NTNH without any of the other OrfX cluster proteins. NTNH subtype-specific immunoreactivity was also discovered, allowing for the differentiation of subtypes based on cluster proteins associated with BoNT.

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