scholarly journals Immobilization of Cells with Surface-Displayed Chitin-Binding Domain

2006 ◽  
Vol 72 (1) ◽  
pp. 927-931 ◽  
Author(s):  
Jen-You Wang ◽  
Yun-Peng Chao
Keyword(s):  
Escherichia Coli ◽  
Gene Fusion ◽  
Cell Immobilization ◽  
Binding Domain ◽  
Engineering Applications ◽  
Chitin Binding Domain ◽  
Wide Range ◽  
Frame Fusion

ABSTRACT To explore chitin-binding domain (ChBD)-based cell immobilization, a tripartite gene fusion consisting of an in-frame fusion of ChBD to lpp and ompA was constructed and expressed in Escherichia coli. ChBD-displayed cells exhibited highly specific and stable binding to chitin within a wide range of pHs (5 to 8) and temperatures (15 to 37°C). These results illustrate the promising use of this approach for engineering applications.

scholarly journals Expression and Characterization of the Chitin-Binding Domain of Chitinase A1 from Bacillus circulans WL-12

2000 ◽  
Vol 182 (11) ◽  
pp. 3045-3054 ◽  
Author(s):  
Masayuki Hashimoto ◽  
Takahisa Ikegami ◽  
Shizuka Seino ◽  
Nobuhumi Ohuchi ◽  
Harumi Fukada ◽  
...  
Keyword(s):  
Hydrophobic Interactions ◽  
Expression System ◽  
Binding Activity ◽  
Binding Domain ◽  
Bacillus Circulans ◽  
Titration Calorimetry ◽  
Affinity Column ◽  
Chitin Binding Domain ◽  
Wide Range ◽  
Hydrolysis Of

ABSTRACT Chitinase A1 from Bacillus circulans WL-12 comprises an N-terminal catalytic domain, two fibronectin type III-like domains, and a C-terminal chitin-binding domain (ChBD). In order to study the biochemical properties and structure of the ChBD, ChBDChiA1 was produced in Escherichia coliusing a pET expression system and purified by chitin affinity column chromatography. Purified ChBDChiA1 specifically bound to various forms of insoluble chitin but not to other polysaccharides, including chitosan, cellulose, and starch. Interaction of soluble chitinous substrates with ChBDChiA1 was not detected by means of nuclear magnetic resonance and isothermal titration calorimetry. In addition, the presence of soluble substrates did not interfere with the binding of ChBDChiA1 to regenerated chitin. These observations suggest that ChBDChiA1recognizes a structure which is present in insoluble or crystalline chitin but not in chito-oligosaccharides or in soluble derivatives of chitin. ChBDChiA1 exhibited binding activity over a wide range of pHs, and the binding activity was enhanced at pHs near its pI and by the presence of NaCl, suggesting that the binding of ChBDChiA1 is mediated mainly by hydrophobic interactions. Hydrolysis of β-chitin microcrystals by intact chitinase A1 and by a deletion derivative lacking the ChBD suggested that the ChBD is not absolutely required for hydrolysis of β-chitin microcrystals but greatly enhances the efficiency of degradation.

scholarly journals Structure of a consensus chitin-binding domain revealed by solution NMR

2020 ◽  
Author(s):  
Dario Heymann ◽  
Harini Mohanram ◽  
Akshita Kumar ◽  
Chandra S. Verma ◽  
Julien Lescar ◽  
...  
Keyword(s):  
Binding Proteins ◽  
3D Structure ◽  
Binding Domain ◽  
Biomimetic Synthesis ◽  
Binding Motif ◽  
Carbohydrate Binding ◽  
Dosidicus Gigas ◽  
Chitin Binding Domain ◽  
Carbohydrate Degradation ◽  
Wide Range

ABSTRACTCarbohydrate-binding proteins (CBPs) are a versatile group of proteins found in almost every organism on earth. CBPs are involved in enzymatic carbohydrate degradation and also serve as templating scaffolds in the exoskeleton of crustaceans and insects. One specific chitin-binding motif found across a wide range of arthropods’ exoskeletons is the “extended Rebers and Riddiford” consensus (R&R). However, how the R&R motif binds chitin is unclear. Here, we report the 3D structure and molecular level interactions of a chitin-binding domain (CBD-γ) located in a CBP from the beak of the jumbo squid Dosidicus gigas. This CBP is one of four chitin-binding proteins identified in the beak mouthpart of D. gigas and is believed to interact with chitin to form a scaffold network that is infiltrated with a second set of structural proteins during beak maturation. We used solution state NMR spectroscopy to elucidate the molecular interactions between CBD-γ and the soluble chitin derivative pentaacetyl-chitopentaose (PCP) and find that folding of this domain is triggered upon its interaction with PCP. To our knowledge, this is the first experimental 3D structure of a CBP containing the R&R consensus motif, which can be used as a template to understand in more details the role of the R&R motif found in a wide range of CBP-chitin complexes. The present structure also provides molecular information for biomimetic synthesis of graded biomaterials using aqueous-based chemistry and biopolymers.

An Improved Membrane Filtration Method for Enumeration of Faecal Coliforms and E. Coli by a Fluorogenic β-Glucuronidase Assay

1993 ◽  
Vol 27 (3-4) ◽  
pp. 267-270 ◽  
Author(s):  
M. T. Augoustinos ◽  
N. A. Grabow ◽  
B. Genthe ◽  
R. Kfir
Keyword(s):  
Escherichia Coli ◽  
Water Samples ◽  
Membrane Filtration ◽  
Faecal Coliforms ◽  
Environmental Waters ◽  
Filtration Method ◽  
E Coli ◽  
Wide Range ◽  
Pure Cultures ◽  
Glucuronidase Assay

A fluorogenic β-glucuronidase assay comprising membrane filtration followed by selective enumeration on m-FC agar at 44.5°C and further confirmation using tlie 4-metliylumbelliferyl-β-D-glucuronide (MUG) containing medium was evaluated for the detection of Escherichia coli in water. A total of 200 typical blue and non-typical blue colonies were isolated from sea and fresh water samples using initial selective enumeration on m-FC agar. Pure cultures of the selected colonies were further tested using the MUG assay and identified using the API 20E method. Of the colonies tested which were shown to be positive using the MUG assay 99.4% were Escherichia coli. The results of this study indicate the combination of the m-FC method followed by the MUG assay to be highly efficient for the selection and confirmation of E. coli from a wide range of environmental waters.

ТЕХНОЛОГИЯ И ПРИМЕНЕНИЕ ЭЛЕКТРОХИМИЧЕСКИХ ПРЕОБРАЗОВАТЕЛЕЙ

2020 ◽  
Vol 96 (3s) ◽  
pp. 450-455
Author(s):  
В.Г. Криштоп ◽  
Д.А. Жевненко ◽  
П.В. Дудкин ◽  
Е.С. Горнев ◽  
В.Г. Попов ◽  
...  
Keyword(s):  
Pressure Sensors ◽  
Engineering Applications ◽  
Electrochemical Systems ◽  
Seismic Sensors ◽  
Microelectronic Technology ◽  
Wide Range ◽  
Electrochemical Transducers ◽  
Element Base ◽  
New Devices

Электрохимические системы очень перспективны для разработки новой элементной базы для микроэлектроники и для использования в широком спектре инженерных задач. Мы разработали новую микроэлектронную технологию для изготовления электрохимических преобразователей (ЭХП) и новые приборы на основе новых электрохимических микроэлектронных чипов. Планарные электрохимические преобразователи могут использоваться в акселерометрах, сейсмических датчиках, датчиках вращения, гидрофонах и датчиках давления. Electrochemical systems are very promising for the development of a new element base for microelectronics, and for use in a wide range of engineering applications. We have developed a new microelectronic technology for manufacturing electrochemical transducers (ECP) and new devices based on new electrochemical microelectronic chips. Planar electrochemical transducers are used in accelerometers, seismic sensors, rotation sensors, hydrophones and pressure sensors.

The lacI Gene as a Target for Mutation in Transgenic Rodents and Escherichia coli

Genetics ◽  
1998 ◽  
Vol 148 (4) ◽  
pp. 1441-1451
Author(s):  
Johan G de Boer ◽  
Barry W Glickman
Keyword(s):  
Escherichia Coli ◽  
Dna Repair ◽  
Dna Binding ◽  
Transgenic Animals ◽  
Single Copy ◽  
Binding Domain ◽  
Dna Binding Domain ◽  
Large Database ◽  
Deletion Mutations ◽  
Laci Gene

Abstract The lacI gene has been used extensively for the recovery and analysis of mutations in bacteria with various DNA repair backgrounds and after exposure to a wide variety of mutagens. This has resulted in a large database of information on mutational mechanisms and specificity of many mutagens, as well as the effect of DNA repair background on mutagenicity. Most importantly, knowledge about the mutational sensitivity of the lacI gene is now available, yielding information about mutable nucleotides. This popularity and available knowledge resulted in the use of the lacI gene in transgenic rodents for the study of mutagenesis in mammals, where it resides in ~40 repeated copies. As the number of sequenced mutations recovered from these animals increases, we are able to analyze the sites at which mutations have been recovered in great detail and to compare the recovered sites between bacteria and transgenic animals. The nucleotides that code for the DNA-binding domain are nearly saturated with base substitutions. Even after determining the sequences of ~10,000 mutations recovered from the animals, however, new sites and new changes are still being recovered. In addition, we compare the nature of deletion mutations between bacteria and animals. Based on the nature of deletions in the animals, we conclude that each deletion occurs in a single copy of the gene.

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