Heterogeneous Expression and Functional Characterization of Cellulose-Degrading Enzymes from Aspergillus niger for Enzymatic Hydrolysis of Alkali Pretreated Bamboo Biomass

2015 ◽  
Vol 57 (9) ◽  
pp. 859-867 ◽  
Author(s):  
Nasir Ali ◽  
Zhang Ting ◽  
Hailong Li ◽  
Yong Xue ◽  
Lihui Gan ◽  
...  
Processes ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 1360
Author(s):  
Ekaterina Budenkova ◽  
Stanislav Sukhikh ◽  
Svetlana Ivanova ◽  
Olga Babich ◽  
Vyacheslav Dolganyuk ◽  
...  

Enzymatic hydrolysis of cellulose-containing raw materials, using Aspergillus niger, were studied. Filter paper, secondary cellulose-containing or starch-containing raw materials, miscanthus cellulose after alkaline or acid pretreatment, and wood chip cellulose, were used as substrates. The study focused on a wild A. niger strain, treated, or not (control), by ultraviolet (UV) irradiations for 45, 60, or 120 min (UV45, UV60, or UV120), or by UV irradiation for 120 min followed by a chemical treatment with NaN3 + ItBr for 30 min or 80 min (UV120 + CH30 or UV120 + CH80). A mixture of all the A. niger strains (MIX) was also tested. A citrate buffer, at 50 mM, wasthe most suitable for enzymatic hydrolysis. As the UV exposure time increased to 2 h, the cellulase activity of the surviving culturewas increased (r = 0.706; p < 0.05). The enzymatic activities of the obtained strains, towards miscanthus cellulose, wood chips, and filter paper, were inferior to those obtained with commercial enzymes (8.6 versus 9.1 IU), in some cases. Under stationary hydrolysis at 37 °C, pH = 4.7, the enzymatic activity of A. niger UV120 + CH30 was 24.9 IU. The enzymatic hydrolysis of secondary raw materials, using treated A. niger strains, was themost effective at 37 °C. Similarly, the most effective treatment of miscanthus cellulose and wood chips occurred at 50 °C. The maximum conversion of cellulose to glucose was observed using miscanthus cellulose (with alkaline pretreatment), and the minimum conversion was observed when using wood chips. The greatest value of cellulase activity was evidenced in the starch-containing raw materials, indicating that A. niger can ferment not only through cellulase activity, but also via an amylolytic one.


2019 ◽  
Vol 8 (3) ◽  
Author(s):  
Alan De Oliveira Campos ◽  
Francinaldo Leite Da Silva ◽  
Emilianny Rafaely Batista Magalhães ◽  
Gorete Ribeiro De Macedo ◽  
Everaldo Silvino Dos Santos

Carnauba (Copernicia prunifera) straw residue generated from production of its wax is rich in cellulose, thus showing a potential use in the production of second generation ethanol. However, the high lignin and hemicellulose load associated with cellulose makes it difficult the enzymatic attack, thus having the need of an adequate pretreatment of this material. The objective of this study was to optimize the enzymatic hydrolysis of carnauba straw residue, focusing on the alkaline biomass pretreatment. Therefore, NaOH solutions at concentrations of 1.0% (w/v) (PA1), 2.0% (w/v) (PA2), 3.0% (w/v) (PA3) and 4.0% (w/v) (PA4) were used. The chemical and physical characterization of natural and pre-treated carnauba straw were according to the NREL, and DRX and FTIR performed analyzes. The materials chemical characterization showed that all the used pretreatments were able to remove a significant amount of lignin and hemicellulose, which can improve the enzymes access, favoring the increase of cellulose conversion. In relation to DRX analysis an increase in crystallinity index happens reaching up to 55.15% after the pretreatment PA4, which may be associated to the removal of hemicellulose and amorphous lignin, related to cellulose. After a period of 96 hours of enzymatic hydrolysis, the PA4 pretreated residue showed the best performance with a cellulosic conversion of 78%. Spite of a slightly lower performance of the residue that presented higher cellulose conversion, the pretreated material PA2 is an alternative to reduce costs in the cellulosic ethanol production.


2005 ◽  
Vol 388 (2) ◽  
pp. 493-500 ◽  
Author(s):  
Chandra N. PATEL ◽  
David W. KOH ◽  
Myron K. JACOBSON ◽  
Marcos A. OLIVEIRA

PARG [poly(ADP-ribose) glycohydrolase] catalyses the hydrolysis of α(1″→2′) or α(1‴→2″) O-glycosidic linkages of ADP-ribose polymers to produce free ADP-ribose. We investigated possible mechanistic similarities between PARG and glycosidases, which also cleave O-glycosidic linkages. Glycosidases typically utilize two acidic residues for catalysis, thus we targeted acidic residues within a conserved region of bovine PARG that has been shown to contain an inhibitor-binding site. The targeted glutamate and aspartate residues were changed to asparagine in order to minimize structural alterations. Mutants were purified and assayed for catalytic activity, as well as binding, to an immobilized PARG inhibitor to determine ability to recognize substrate. Our investigation revealed residues essential for PARG catalytic activity. Two adjacent glutamic acid residues are found in the conserved sequence Gln755-Glu-Glu757, and a third residue found in the conserved sequence Val737-Asp-Phe-Ala-Asn741. Our functional characterization of PARG residues, along with recent identification of an inhibitor-binding residue Tyr796 and a glycine-rich region Gly745-Gly-Gly747 important for PARG function, allowed us to define a PARG ‘signature sequence’ [vDFA-X3-GGg-X6–8-vQEEIRF-X3-PE-X14-E-X12-YTGYa], which we used to identify putative PARG sequences across a range of organisms. Sequence alignments, along with our mapping of PARG functional residues, suggest the presence of a conserved catalytic domain of approx. 185 residues which spans residues 610–795 in bovine PARG.


2020 ◽  
Vol 14 (4) ◽  
pp. 453-460
Author(s):  
Ruyi Sha ◽  
Zhan Yu ◽  
Zhenzhen Wang ◽  
Edwin Menledy Gbor ◽  
Ligang Jiang ◽  
...  

The lignin present in lignocellulose seriously affects the efficiency of cellulose enzymatic hydrolysis. In addition, lignin adsorbs high-cost cellulase, causing greater economic losses. Lignin can also disturb the site of action of cellulase and reduce the efficiency of hydrolysis. Therefore, if lignin is removed or surface modified before cellulose enzymatic hydrolysis, the enzymatic hydrolysis efficiency of lignocellulosic biomass will be greatly improved. In this paper, the cellulose enzymatic properties of bamboo biomass being treated with dilute acid and alkaline under the intervention of biosurfactant rhamnolipid were evaluated. The effects of rhamnolipids on the adsorption characterization of cellulose on pretreated bamboo were studied. Besides, the inter-communication between rhamnolipids and cellulose was investigated by fluorescence probe. The results showed that rhamnolipids could have a positive effect on the enzymatic hydrolysis of bamboo biomass by reducing the non-productive adsorption of cellulase on the surface of lignocellulose. The outcome illustrated that cellulase could be combined with rhamnolipids micelles, participating in the formation of rhamnolipids micelles, thereby increasing the internal hydrophobicity of the micelles, but could not change the properties of rhamnolipids micelles higher than one CMC (Critical Micelle Concentration). It can be seen that the interaction between rhamnolipids and cellulase is beneficial to enhance the stability and enzymatic activity of cellulase, thereby improving the enzymatic hydrolysis efficiency of cellulose in biomass. Based on these results, a theoretical knowledge about the mechanism of enhancing the enzymatic hydrolysis efficiency of lignocellulose by biosurfactants rhamnolipids is provided.


BioResources ◽  
2016 ◽  
Vol 12 (1) ◽  
Author(s):  
Yin Ying H'ng ◽  
Akiko Nakagawa-Izumi ◽  
Cheu Peng Leh ◽  
Atanu Kumar Das ◽  
Hiroshi Ohi

1994 ◽  
Vol 302 (2) ◽  
pp. 455-461 ◽  
Author(s):  
R M Goraczniak ◽  
T Duda ◽  
A Sitaramayya ◽  
R K Sharma

In the vertebrate photoreceptor cell, rod outer segment (ROS) is the site of visual signal-transduction process, and a pivotal molecule that regulates this process is cyclic GMP. Cyclic GMP controls the cationic conductance into the ROS, and light causes a decrease in the conductance by activating hydrolysis of the cyclic nucleotide. The identity of the granylate cyclase (ROS-GC) that synthesizes this pool of cyclic GMP is unknown. We now report the cloning, expression and functional characterization of a DNA from bovine retina that encodes ROS-GC.


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