A simple and nondestructive method for the separation of polysaccharides from β-glucosidase produced extracellularly by Aspergillus niger

Khawar Sohail Siddiqui; Muhammad Hamid Rashid; Ahsan Mushir Shemsi; Muhammad Ibrahim Rajoka

doi:10.1016/0141-0229(94)90067-1

A simple and nondestructive method for the separation of polysaccharides from β-glucosidase produced extracellularly by Aspergillus niger

Khawar Sohail Siddiqui^*, Muhammad Hamid Rashid, Ahsan Mushir Shemsi, Muhammad Ibrahim Rajoka

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

An apparatus based on electrophoresis has been devised that removes noncovalently bound polysaccharides from extracellular proteins of Aspergillus niger with concomitant partial β-glucosidase purification and concentration. The apparatus consists of a series of three chambers separated by polyacrylamide gels. Dialyzed and concentrated crude extract of Aspergillus niger containing β-glucosidase was poured into the middle chamber, while smaller anodic and cathodic chambers contained buffer. When electric current was applied, negatively charged protein-polysaccharide complexes moved toward the anode. Most of the negatively charged proteins, including β-glucosidase, crossed the gel barrier into the anodic compartment, while neutral polysaccharides were either trapped in the gels or remained in the middle chamber. In this way, 125 ml of dialyzed and concentrated crude extract of Aspergillus niger was processed. Therefore, after 24 h of electrophoresis, 68% of the proteins and 90% of the β-glucosidase activity, but only negligible amounts of polysaccharide, were transferred to the anodic chamber. The removal of high-molecular-weight polysaccharide from β-glucosidase had a detrimental effect on the stability of the enzyme.

Original language	English
Pages (from-to)	912-917
Number of pages	6
Journal	Enzyme and Microbial Technology
Volume	16
Issue number	10
DOIs	https://doi.org/10.1016/0141-0229(94)90067-1
State	Published - Oct 1994
Externally published	Yes

Bibliographical note

Funding Information:
The work described in this paper is part of the Ph.D. research of M. H. Rashid. This work was financed in part by a grant made by the United States Agency for International Development under PSTC proposal 6-163, USAID grant No. 9365542-G-00-89-42-00, and PAEC. We wish to acknowledge Director NIBGE, Dr. K. A. Malik, for providing research facilities. We would also like to thank M. H. Rash-id's external supervisor, Prof. A. R. Shakoori, for his valuable suggestions. We are thankful to Mr. Tanvir Ahmad for his help in computer graphics. The technical assistance of G. A. Waseer is appreciated.

Keywords

Aspergillus niger
Polysaccharides
compartmental electrophoresis
β-glucosidase

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biochemistry
Applied Microbiology and Biotechnology

Access to Document

10.1016/0141-0229(94)90067-1

Cite this

@article{e230b3c2a2b348fbaacbc78fb4f3b890,

title = "A simple and nondestructive method for the separation of polysaccharides from β-glucosidase produced extracellularly by Aspergillus niger",

abstract = "An apparatus based on electrophoresis has been devised that removes noncovalently bound polysaccharides from extracellular proteins of Aspergillus niger with concomitant partial β-glucosidase purification and concentration. The apparatus consists of a series of three chambers separated by polyacrylamide gels. Dialyzed and concentrated crude extract of Aspergillus niger containing β-glucosidase was poured into the middle chamber, while smaller anodic and cathodic chambers contained buffer. When electric current was applied, negatively charged protein-polysaccharide complexes moved toward the anode. Most of the negatively charged proteins, including β-glucosidase, crossed the gel barrier into the anodic compartment, while neutral polysaccharides were either trapped in the gels or remained in the middle chamber. In this way, 125 ml of dialyzed and concentrated crude extract of Aspergillus niger was processed. Therefore, after 24 h of electrophoresis, 68\% of the proteins and 90\% of the β-glucosidase activity, but only negligible amounts of polysaccharide, were transferred to the anodic chamber. The removal of high-molecular-weight polysaccharide from β-glucosidase had a detrimental effect on the stability of the enzyme.",

keywords = "Aspergillus niger, Polysaccharides, compartmental electrophoresis, β-glucosidase",

author = "Siddiqui, \{Khawar Sohail\} and Rashid, \{Muhammad Hamid\} and Shemsi, \{Ahsan Mushir\} and Rajoka, \{Muhammad Ibrahim\}",

note = "Funding Information: The work described in this paper is part of the Ph.D. research of M. H. Rashid. This work was financed in part by a grant made by the United States Agency for International Development under PSTC proposal 6-163, USAID grant No. 9365542-G-00-89-42-00, and PAEC. We wish to acknowledge Director NIBGE, Dr. K. A. Malik, for providing research facilities. We would also like to thank M. H. Rash-id's external supervisor, Prof. A. R. Shakoori, for his valuable suggestions. We are thankful to Mr. Tanvir Ahmad for his help in computer graphics. The technical assistance of G. A. Waseer is appreciated.",

year = "1994",

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doi = "10.1016/0141-0229(94)90067-1",

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journal = "Enzyme and Microbial Technology",

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AU - Rashid, Muhammad Hamid

AU - Shemsi, Ahsan Mushir

AU - Rajoka, Muhammad Ibrahim

N1 - Funding Information: The work described in this paper is part of the Ph.D. research of M. H. Rashid. This work was financed in part by a grant made by the United States Agency for International Development under PSTC proposal 6-163, USAID grant No. 9365542-G-00-89-42-00, and PAEC. We wish to acknowledge Director NIBGE, Dr. K. A. Malik, for providing research facilities. We would also like to thank M. H. Rash-id's external supervisor, Prof. A. R. Shakoori, for his valuable suggestions. We are thankful to Mr. Tanvir Ahmad for his help in computer graphics. The technical assistance of G. A. Waseer is appreciated.

PY - 1994/10

Y1 - 1994/10

N2 - An apparatus based on electrophoresis has been devised that removes noncovalently bound polysaccharides from extracellular proteins of Aspergillus niger with concomitant partial β-glucosidase purification and concentration. The apparatus consists of a series of three chambers separated by polyacrylamide gels. Dialyzed and concentrated crude extract of Aspergillus niger containing β-glucosidase was poured into the middle chamber, while smaller anodic and cathodic chambers contained buffer. When electric current was applied, negatively charged protein-polysaccharide complexes moved toward the anode. Most of the negatively charged proteins, including β-glucosidase, crossed the gel barrier into the anodic compartment, while neutral polysaccharides were either trapped in the gels or remained in the middle chamber. In this way, 125 ml of dialyzed and concentrated crude extract of Aspergillus niger was processed. Therefore, after 24 h of electrophoresis, 68% of the proteins and 90% of the β-glucosidase activity, but only negligible amounts of polysaccharide, were transferred to the anodic chamber. The removal of high-molecular-weight polysaccharide from β-glucosidase had a detrimental effect on the stability of the enzyme.

AB - An apparatus based on electrophoresis has been devised that removes noncovalently bound polysaccharides from extracellular proteins of Aspergillus niger with concomitant partial β-glucosidase purification and concentration. The apparatus consists of a series of three chambers separated by polyacrylamide gels. Dialyzed and concentrated crude extract of Aspergillus niger containing β-glucosidase was poured into the middle chamber, while smaller anodic and cathodic chambers contained buffer. When electric current was applied, negatively charged protein-polysaccharide complexes moved toward the anode. Most of the negatively charged proteins, including β-glucosidase, crossed the gel barrier into the anodic compartment, while neutral polysaccharides were either trapped in the gels or remained in the middle chamber. In this way, 125 ml of dialyzed and concentrated crude extract of Aspergillus niger was processed. Therefore, after 24 h of electrophoresis, 68% of the proteins and 90% of the β-glucosidase activity, but only negligible amounts of polysaccharide, were transferred to the anodic chamber. The removal of high-molecular-weight polysaccharide from β-glucosidase had a detrimental effect on the stability of the enzyme.

KW - Aspergillus niger

KW - Polysaccharides

KW - compartmental electrophoresis

KW - β-glucosidase

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JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

IS - 10

ER -

A simple and nondestructive method for the separation of polysaccharides from β-glucosidase produced extracellularly by Aspergillus niger

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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