Effect of LPMO on the Hydrolysis of Crystalline Chitin by Chitinase A and β-N-Acetylglucosaminidase from Paenibacillus sp.

2020 ◽  
Author(s):  
Mitsuhiro Ueda ◽  
Kei Nakadoi ◽  
Kana Tsukamoto ◽  
Shunsuke Sakurai
Keyword(s):  
Chitinase A ◽  
Paenibacillus Sp ◽  
Hydrolysis Of

scholarly journals Production of Thermophilic Chitinase by Paenibacillus sp. TKU052 by Bioprocessing of Chitinous Fishery Wastes and Its Application in N-acetyl-D-glucosamine Production

Polymers ◽  
2021 ◽  
Vol 13 (18) ◽  
pp. 3048
Author(s):  
Chien Thang Doan ◽  
Thi Ngoc Tran ◽  
San-Lang Wang
Keyword(s):  
Enzyme Production ◽  
Production Cost ◽  
Maximum Activity ◽  
Paenibacillus Sp ◽  
Negative Effect ◽  
Chitinase Production ◽  
Tween 40 ◽  
Triton X ◽  
Catalytic Functions ◽  
Hydrolysis Of

The bioprocessing of chitinous fishery wastes (CFWs) to chitinases through fermentation approaches has gained importance owing to its great benefits in reducing the enzyme production cost, and utilizing chitin waste. In this work, our study of the chitinase production of Paenibacillus sp. TKU052 in the presence of different kinds of CFWs revealed a preference for demineralized crab shells powder (deCSP); furthermore, a 72 kDa chitinase was isolated from the 0.5% deCSP-containing medium. The Paenibacillus sp. TKU052 chitinase displayed maximum activity at 70 °C and pH 4–5, while Zn2+, Fe3+, Triton X-100, Tween 40, and SDS exerted a negative effect on its activity, whereas Mn2+ and 2-mercaptoethanol were found to potentially enhance the activity. Among various kinds of polysaccharide, Paenibacillus sp. TKU052 chitinase exhibited the best catalytic activity on colloidal chitin (CC) with Km = 9.75 mg/mL and Vmax = 2.43 μmol/min. The assessment of the hydrolysis of CC and N-acetyl chitooligosaccharides revealed that Paenibacillus sp. TKU052 chitinase possesses multiple catalytic functions, including exochitinase, endochitinase, and N-acetyl-β-D-glucosaminidase activities. Finally, the combination of Paenibacillus sp. TKU052 chitinase and Streptomyces speibonae TKU048 N-acetyl-β-D-glucosaminidase could efficiently convert CC to N-acetyl-D-glucosamine (GlcNAc) with a production yield of 94.35–98.60% in 12–24 h.

scholarly journals Family 18 chitinase–oligosaccharide substrate interaction: subsite preference and anomer selectivity of Serratia marcescens chitinase A

2003 ◽  
Vol 376 (1) ◽  
pp. 87-95 ◽  
Author(s):  
Nathan N. ARONSON ◽  
Brian A. HALLORAN ◽  
Mikhail F. ALEXYEV ◽  
Lauren AMABLE ◽  
Jeffry D. MADURA ◽  
...  
Keyword(s):  
Serratia Marcescens ◽  
Substrate Binding ◽  
Binding Mode ◽  
Wild Type ◽  
Chitinase A ◽  
Novel Approach ◽  
The Family ◽  
Binding Cleft ◽  
Hydrolysis Of ◽  
Site Binding

The sizes and anomers of the products formed during the hydrolysis of chitin oligosaccharides by the Family 18 chitinase A (ChiA) from Serratia marcescens were analysed by hydrophilic interaction chromatography using a novel approach in which reactions were performed at 0 °C to stabilize the anomer conformations of the initial products. Crystallographic studies of the enzyme, having the structure of the complex of the ChiA E315L (Glu315→Leu) mutant with a hexasaccharide, show that the oligosaccharide occupies subsites −4 to +2 in the substrate-binding cleft, consistent with the processing of β-chitin by the release of disaccharide at the reducing end. Products of the hydrolysis of hexa- and penta-saccharides by wild-type ChiA, as well as by two mutants of the residues Trp275 and Phe396 important in binding the substrate at the +1 and +2 sites, show that the substrates only occupy sites −2 to +2 and that additional N-acetyl-d-glucosamines extend beyond the substrate-binding cleft at the reducing end. The subsites −3 and −4 are not used in this four-site binding mode. The explanation for these results is found in the high importance of individual binding sites for the processing of short oligosaccharides compared with the cumulative recognition and processive hydrolysis mechanism used to digest natural β-chitin.

scholarly journals Effective valorization of barley bran for simultaneous cellulase and β-amylase production by Paenibacillus chitinolyticus CKS1: Statistical optimization and enzymes application

2017 ◽  
Vol 82 (11) ◽  
pp. 1223-1236 ◽  
Author(s):  
Katarina Mihajlovski ◽  
Sladjana Davidovic ◽  
Djordje Veljovic ◽  
Milica Carevic ◽  
Vesna Lazic ◽  
...  
Keyword(s):  
High Performance ◽  
Raw Materials ◽  
Starch Structure ◽  
Cotton Fibres ◽  
Hydrolysis Process ◽  
Paenibacillus Sp ◽  
Amylase Production ◽  
Barley Bran ◽  
Hydrolysis Of ◽  
By Products

The agricultural raw industry generates large amounts of annually by- -products that create disposal problems. Hitherto, there have been no reported papers about the simultaneous production of cellulase and ?-amylase from these raw materials using Paenibacillus sp. that would reduce the costs. Thus, in this paper simultaneous cellulase (CMC-ase and avicelase) and ?-amylase production using barley bran and the application of the natural isolate Paenibacillus chitinolyticus CKS1 and potential enzymes in the hydrolysis process was studied. Response surface methodology was used to obtain the maximum enzyme activity (CMC-ase 0.405 U mL-1, avicelase 0.433 U mL-1 and ?-amylase 1.594U mL-1). Scanning electron microscopy showed degradation of the lignocellulosic?starch structure of barley bran after fermentation. The CKS1 bacterial supernatant, which contains cellulases and ?-amylase, could hydrolyze cotton fibres and barley bran, respectively. The main products after enzymatic hydrolysis of cotton fibres and barley bran, glucose (0.117 g gmat -1) and maltose (0.347 g gmat -1), were quantified by high performance liquid chromatography (HPLC). The produced enzymes could be used for hydrolysis of cotton fabric and barley bran to glucose and maltose, respectively. Application of simultaneous enzymes production using an agricultural by-product is economically and environmentally accepted and moreover, valuable biotechnological products, such as glucose and maltose, were obtained in this investigation.

Fine Structural Localization of Acyltransferases Involved in Lipid Synthesis in Intestinal Mucosa.

1970 ◽  
Vol 28 ◽  
pp. 54-55
Author(s):  
R. J. Barrnett ◽  
J. A. Higgins
Keyword(s):  
Smooth Endoplasmic Reticulum ◽  
Lipid Synthesis ◽  
Mucosal Cell ◽  
Structural Localization ◽  
Morphological Studies ◽  
Mucosal Cells ◽  
Initial Appearance ◽  
Sh Group ◽  
Hydrolysis Of ◽  
Intestinal Mucosal Cells

The main products of intestinal hydrolysis of dietary triglycerides are free fatty acids and monoglycerides. These form micelles from which the lipids are absorbed across the mucosal cell brush border. Biochemical studies have indicated that intestinal mucosal cells possess a triglyceride synthesising system, which uses monoglyceride directly as an acylacceptor as well as the system found in other tissues in which alphaglycerophosphate is the acylacceptor. The former pathway is used preferentially for the resynthesis of triglyceride from absorbed lipid, while the latter is used mainly for phospholipid synthesis. Both lipids are incorporated into chylomicrons. Morphological studies have shown that during fat absorption there is an initial appearance of fat droplets within the cisternae of the smooth endoplasmic reticulum and that these subsequently accumulate in the golgi elements from which they are released at the lateral borders of the cell as chylomicrons.We have recently developed several methods for the fine structural localization of acyltransferases dependent on the precipitation, in an electron dense form, of CoA released during the transfer of the acyl group to an acceptor, and have now applied these methods to a study of the fine structural localization of the enzymes involved in chylomicron lipid biosynthesis. These methods are based on the reduction of ferricyanide ions by the free SH group of CoA.

Lattice imaging and pore structure of β ferric oxyhydroxide

1978 ◽  
Vol 36 (1) ◽  
pp. 264-265
Author(s):  
T. Baird ◽  
J.R. Fryer ◽  
S.T. Galbraith
Keyword(s):  
Electron Microscopy ◽  
Room Temperature ◽  
Bulk Material ◽  
Model Structure ◽  
Bulk Crystals ◽  
Lattice Imaging ◽  
Thin Sections ◽  
Ferric Oxyhydroxide ◽  
Resolution Electron ◽  
Hydrolysis Of

Introduction Previously we had suggested (l) that the striations observed in the pod shaped crystals of β FeOOH were an artefact of imaging in the electron microscope. Contrary to this adsorption measurements on bulk material had indicated the presence of some porosity and Gallagher (2) had proposed a model structure - based on the hollandite structure - showing the hollandite rods forming the sides of 30Å pores running the length of the crystal. Low resolution electron microscopy by Watson (3) on sectioned crystals embedded in methylmethacrylate had tended to support the existence of such pores.We have applied modern high resolution techniques to the bulk crystals and thin sections of them without confirming these earlier postulatesExperimental β FeOOH was prepared by room temperature hydrolysis of 0.01M solutions of FeCl3.6H2O, The precipitate was washed, dried in air, and embedded in Scandiplast resin. The sections were out on an LKB III Ultramicrotome to a thickness of about 500Å.

Elucidating cyclic AMP signaling in subcellular domains with optogenetic tools and fluorescent biosensors

2019 ◽  
Vol 47 (6) ◽  
pp. 1733-1747 ◽  
Author(s):  
Christina Klausen ◽  
Fabian Kaiser ◽  
Birthe Stüven ◽  
Jan N. Hansen ◽  
Dagmar Wachten
Keyword(s):  
Cyclic Amp ◽  
Adenosine Monophosphate ◽  
Adenylyl Cyclases ◽  
Temporal Precision ◽  
Fluorescent Biosensors ◽  
Subcellular Domains ◽  
Spatio Temporal ◽  
Hydrolysis Of ◽  
Shed Light ◽  
Cyclic Amp Signaling

The second messenger 3′,5′-cyclic nucleoside adenosine monophosphate (cAMP) plays a key role in signal transduction across prokaryotes and eukaryotes. Cyclic AMP signaling is compartmentalized into microdomains to fulfil specific functions. To define the function of cAMP within these microdomains, signaling needs to be analyzed with spatio-temporal precision. To this end, optogenetic approaches and genetically encoded fluorescent biosensors are particularly well suited. Synthesis and hydrolysis of cAMP can be directly manipulated by photoactivated adenylyl cyclases (PACs) and light-regulated phosphodiesterases (PDEs), respectively. In addition, many biosensors have been designed to spatially and temporarily resolve cAMP dynamics in the cell. This review provides an overview about optogenetic tools and biosensors to shed light on the subcellular organization of cAMP signaling.

Sign in / Sign up

Export Citation Format

Share Document