Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D

Qiang Li, Linda Harvey, Brian McNeil

Research output: Contribution to conferencePoster

Abstract

Conference poster illustrating how filamentous fungi have attracted extensive research interest due to their abilities to secrete large amounts of high-valued recombinant proteins. Despite the fact that proteases have been recognized as one of the main problems in protein production, little is known about the regulation of them. In the present study, we hypothesized that reactive oxygen species (ROS), unavoidable by-products in all aerobic cultures, may cause protein oxidation and induce proteolysis in filamentous fungi. To test this hypothesis, we used a variety of oxidative stressors, oxygenation, menadione, hydrogen peroxide, hydrogen peroxide with copper ion, to study the induction of intracellular proteolytic activities in A. niger B1-D, a recombinant filamentous fungus secreting hen egg white lysozyme. Protein carbonyl content was monitored as a bio-marker for protein oxidation. Our results show that oxygenation and metal-catalyzed oxidation significantly induce carbonyl groups to bovine serum albumin (BSA). We also found that proteolytic activities and carbonyl content increase on the addition of these stressors to the whole broth in A. niger B1-D, and the responses are dose-dependent. In conclusion, ROS overproduction correlates with protein oxidation and proteolysis in A. niger B1-D. It is advisable to decrease ROS production in the bioprocess of filamentous fungi in order to achieve a higher productivity of heterologous proteins.

Conference

ConferenceFourth Recombinant Protein Production Meeting: a comparative view on host physiology
CityBarcelona, Spain
Period21/09/0623/09/06

Fingerprint

Aspergillus niger
oxidation
fungi
degradation
reactive oxygen species
proteins
proteolysis
hydrogen peroxide
menadione
egg albumen
bovine serum albumin
lysozyme
recombinant proteins
byproducts
hens
proteinases
copper
metals
ions
dosage

Keywords

  • poster
  • protein oxidation
  • microbial cells
  • Aspergillus niger
  • applied chemistry

Cite this

Li, Q., Harvey, L., & McNeil, B. (2006). Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D. 6-6. Poster session presented at Fourth Recombinant Protein Production Meeting: a comparative view on host physiology, Barcelona, Spain, . https://doi.org/10.1186/1475-2859-5-S1-P6
Li, Qiang ; Harvey, Linda ; McNeil, Brian. / Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D. Poster session presented at Fourth Recombinant Protein Production Meeting: a comparative view on host physiology, Barcelona, Spain, .
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abstract = "Conference poster illustrating how filamentous fungi have attracted extensive research interest due to their abilities to secrete large amounts of high-valued recombinant proteins. Despite the fact that proteases have been recognized as one of the main problems in protein production, little is known about the regulation of them. In the present study, we hypothesized that reactive oxygen species (ROS), unavoidable by-products in all aerobic cultures, may cause protein oxidation and induce proteolysis in filamentous fungi. To test this hypothesis, we used a variety of oxidative stressors, oxygenation, menadione, hydrogen peroxide, hydrogen peroxide with copper ion, to study the induction of intracellular proteolytic activities in A. niger B1-D, a recombinant filamentous fungus secreting hen egg white lysozyme. Protein carbonyl content was monitored as a bio-marker for protein oxidation. Our results show that oxygenation and metal-catalyzed oxidation significantly induce carbonyl groups to bovine serum albumin (BSA). We also found that proteolytic activities and carbonyl content increase on the addition of these stressors to the whole broth in A. niger B1-D, and the responses are dose-dependent. In conclusion, ROS overproduction correlates with protein oxidation and proteolysis in A. niger B1-D. It is advisable to decrease ROS production in the bioprocess of filamentous fungi in order to achieve a higher productivity of heterologous proteins.",
keywords = "poster, protein oxidation, microbial cells, Aspergillus niger, applied chemistry",
author = "Qiang Li and Linda Harvey and Brian McNeil",
note = "The 4th Recombinant Protein Production Meeting: a comparative view on host physiology Barcelona, Spain. 21-23 September 2006; Fourth Recombinant Protein Production Meeting: a comparative view on host physiology ; Conference date: 21-09-2006 Through 23-09-2006",
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Li, Q, Harvey, L & McNeil, B 2006, 'Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D' Fourth Recombinant Protein Production Meeting: a comparative view on host physiology, Barcelona, Spain, 21/09/06 - 23/09/06, pp. 6-6. https://doi.org/10.1186/1475-2859-5-S1-P6

Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D. / Li, Qiang; Harvey, Linda; McNeil, Brian.

2006. 6-6 Poster session presented at Fourth Recombinant Protein Production Meeting: a comparative view on host physiology, Barcelona, Spain, .

Research output: Contribution to conferencePoster

TY - CONF

T1 - Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D

AU - Li, Qiang

AU - Harvey, Linda

AU - McNeil, Brian

N1 - The 4th Recombinant Protein Production Meeting: a comparative view on host physiology Barcelona, Spain. 21-23 September 2006

PY - 2006

Y1 - 2006

N2 - Conference poster illustrating how filamentous fungi have attracted extensive research interest due to their abilities to secrete large amounts of high-valued recombinant proteins. Despite the fact that proteases have been recognized as one of the main problems in protein production, little is known about the regulation of them. In the present study, we hypothesized that reactive oxygen species (ROS), unavoidable by-products in all aerobic cultures, may cause protein oxidation and induce proteolysis in filamentous fungi. To test this hypothesis, we used a variety of oxidative stressors, oxygenation, menadione, hydrogen peroxide, hydrogen peroxide with copper ion, to study the induction of intracellular proteolytic activities in A. niger B1-D, a recombinant filamentous fungus secreting hen egg white lysozyme. Protein carbonyl content was monitored as a bio-marker for protein oxidation. Our results show that oxygenation and metal-catalyzed oxidation significantly induce carbonyl groups to bovine serum albumin (BSA). We also found that proteolytic activities and carbonyl content increase on the addition of these stressors to the whole broth in A. niger B1-D, and the responses are dose-dependent. In conclusion, ROS overproduction correlates with protein oxidation and proteolysis in A. niger B1-D. It is advisable to decrease ROS production in the bioprocess of filamentous fungi in order to achieve a higher productivity of heterologous proteins.

AB - Conference poster illustrating how filamentous fungi have attracted extensive research interest due to their abilities to secrete large amounts of high-valued recombinant proteins. Despite the fact that proteases have been recognized as one of the main problems in protein production, little is known about the regulation of them. In the present study, we hypothesized that reactive oxygen species (ROS), unavoidable by-products in all aerobic cultures, may cause protein oxidation and induce proteolysis in filamentous fungi. To test this hypothesis, we used a variety of oxidative stressors, oxygenation, menadione, hydrogen peroxide, hydrogen peroxide with copper ion, to study the induction of intracellular proteolytic activities in A. niger B1-D, a recombinant filamentous fungus secreting hen egg white lysozyme. Protein carbonyl content was monitored as a bio-marker for protein oxidation. Our results show that oxygenation and metal-catalyzed oxidation significantly induce carbonyl groups to bovine serum albumin (BSA). We also found that proteolytic activities and carbonyl content increase on the addition of these stressors to the whole broth in A. niger B1-D, and the responses are dose-dependent. In conclusion, ROS overproduction correlates with protein oxidation and proteolysis in A. niger B1-D. It is advisable to decrease ROS production in the bioprocess of filamentous fungi in order to achieve a higher productivity of heterologous proteins.

KW - poster

KW - protein oxidation

KW - microbial cells

KW - Aspergillus niger

KW - applied chemistry

UR - http://www.biomedcentral.com/content/pdf/1475-2859-5-S1-info.pdf

UR - http://dx.doi.org/10.1186/1475-2859-5-S1-P6

U2 - 10.1186/1475-2859-5-S1-P6

DO - 10.1186/1475-2859-5-S1-P6

M3 - Poster

SP - 6

EP - 6

ER -

Li Q, Harvey L, McNeil B. Native and heterologous protein oxidation and subsequent degradation in a recombinant filamentous fungus Aspergillus niger B1-D. 2006. Poster session presented at Fourth Recombinant Protein Production Meeting: a comparative view on host physiology, Barcelona, Spain, . https://doi.org/10.1186/1475-2859-5-S1-P6