Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus

Barry Murphy; Katie Anderson; Charles Borissow; Patrick Caffrey; Gerald Griffith; Jessica Hearn; Odubunmi Ibrahim; Naseem Khan; Natalie Lamburn; Michael Lee; Katherine Pugh; Bernard Rawlings

doi:10.1039/b922074g

Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus

Barry Murphy, Katie Anderson, Charles Borissow, Patrick Caffrey, Gerald Griffith, Jessica Hearn, Odubunmi Ibrahim, Naseem Khan, Natalie Lamburn, Michael Lee, Katherine Pugh, Bernard Rawlings

Biomedical Sciences

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

Abstract

Amphotericin B is a powerful but toxic drug used against fungal infections and leishmaniases. These diseases would be treated more effectively if non-toxic amphotericin derivatives could be produced on a large scale at low cost. Genetic manipulation of the amphotericin B producer, Streptomyces nodosus, has previously led to the detection and partial characterisation of 8-deoxyamphotericin B, 16-descarboxyl-16-methyl-amphotericin B, 15-deoxy-16-descarboxyl-16-methyl-15-oxo-amphotericin B, 7-oxo-amphotericin B and pentaene analogues. Here we report improved production and purification protocols that have allowed detailed chemical analyses of these compounds. The polyketide synthase product 8-deoxy-16-descarboxyl-16-methyl-amphoteronolide B was identified for the first time. In addition, the ketoreductase 10 domain of the polyketide synthase was specifically inactivated by targeted gene replacement. The resulting mutants produced truncated polyketide intermediates as linear polyenyl-pyrones.

Original language	English
Pages (from-to)	3758-70
Number of pages	13
Journal	Organic and Biomolecular Chemistry
Volume	8
Issue number	16
DOIs	https://doi.org/10.1039/b922074g
Publication status	Published - 21 Aug 2010

Keywords

Amphotericin B/analogs & derivatives
Bacterial Proteins/genetics
Molecular Structure
Protein Engineering
Streptomyces/chemistry

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10.1039/b922074g

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Murphy, B., Anderson, K., Borissow, C., Caffrey, P., Griffith, G., Hearn, J., Ibrahim, O., Khan, N., Lamburn, N., Lee, M., Pugh, K., & Rawlings, B. (2010). Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus. Organic and Biomolecular Chemistry, 8(16), 3758-70. https://doi.org/10.1039/b922074g

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title = "Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus",

abstract = "Amphotericin B is a powerful but toxic drug used against fungal infections and leishmaniases. These diseases would be treated more effectively if non-toxic amphotericin derivatives could be produced on a large scale at low cost. Genetic manipulation of the amphotericin B producer, Streptomyces nodosus, has previously led to the detection and partial characterisation of 8-deoxyamphotericin B, 16-descarboxyl-16-methyl-amphotericin B, 15-deoxy-16-descarboxyl-16-methyl-15-oxo-amphotericin B, 7-oxo-amphotericin B and pentaene analogues. Here we report improved production and purification protocols that have allowed detailed chemical analyses of these compounds. The polyketide synthase product 8-deoxy-16-descarboxyl-16-methyl-amphoteronolide B was identified for the first time. In addition, the ketoreductase 10 domain of the polyketide synthase was specifically inactivated by targeted gene replacement. The resulting mutants produced truncated polyketide intermediates as linear polyenyl-pyrones.",

keywords = "Amphotericin B/analogs & derivatives, Bacterial Proteins/genetics, Molecular Structure, Protein Engineering, Streptomyces/chemistry",

author = "Barry Murphy and Katie Anderson and Charles Borissow and Patrick Caffrey and Gerald Griffith and Jessica Hearn and Odubunmi Ibrahim and Naseem Khan and Natalie Lamburn and Michael Lee and Katherine Pugh and Bernard Rawlings",

year = "2010",

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doi = "10.1039/b922074g",

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Murphy, B, Anderson, K, Borissow, C, Caffrey, P, Griffith, G, Hearn, J, Ibrahim, O, Khan, N, Lamburn, N, Lee, M, Pugh, K & Rawlings, B 2010, 'Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus', Organic and Biomolecular Chemistry, vol. 8, no. 16, pp. 3758-70. https://doi.org/10.1039/b922074g

TY - JOUR

T1 - Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus

AU - Murphy, Barry

AU - Anderson, Katie

AU - Borissow, Charles

AU - Caffrey, Patrick

AU - Griffith, Gerald

AU - Hearn, Jessica

AU - Ibrahim, Odubunmi

AU - Khan, Naseem

AU - Lamburn, Natalie

AU - Lee, Michael

AU - Pugh, Katherine

AU - Rawlings, Bernard

PY - 2010/8/21

Y1 - 2010/8/21

N2 - Amphotericin B is a powerful but toxic drug used against fungal infections and leishmaniases. These diseases would be treated more effectively if non-toxic amphotericin derivatives could be produced on a large scale at low cost. Genetic manipulation of the amphotericin B producer, Streptomyces nodosus, has previously led to the detection and partial characterisation of 8-deoxyamphotericin B, 16-descarboxyl-16-methyl-amphotericin B, 15-deoxy-16-descarboxyl-16-methyl-15-oxo-amphotericin B, 7-oxo-amphotericin B and pentaene analogues. Here we report improved production and purification protocols that have allowed detailed chemical analyses of these compounds. The polyketide synthase product 8-deoxy-16-descarboxyl-16-methyl-amphoteronolide B was identified for the first time. In addition, the ketoreductase 10 domain of the polyketide synthase was specifically inactivated by targeted gene replacement. The resulting mutants produced truncated polyketide intermediates as linear polyenyl-pyrones.

AB - Amphotericin B is a powerful but toxic drug used against fungal infections and leishmaniases. These diseases would be treated more effectively if non-toxic amphotericin derivatives could be produced on a large scale at low cost. Genetic manipulation of the amphotericin B producer, Streptomyces nodosus, has previously led to the detection and partial characterisation of 8-deoxyamphotericin B, 16-descarboxyl-16-methyl-amphotericin B, 15-deoxy-16-descarboxyl-16-methyl-15-oxo-amphotericin B, 7-oxo-amphotericin B and pentaene analogues. Here we report improved production and purification protocols that have allowed detailed chemical analyses of these compounds. The polyketide synthase product 8-deoxy-16-descarboxyl-16-methyl-amphoteronolide B was identified for the first time. In addition, the ketoreductase 10 domain of the polyketide synthase was specifically inactivated by targeted gene replacement. The resulting mutants produced truncated polyketide intermediates as linear polyenyl-pyrones.

KW - Amphotericin B/analogs & derivatives

KW - Bacterial Proteins/genetics

KW - Molecular Structure

KW - Protein Engineering

KW - Streptomyces/chemistry

U2 - 10.1039/b922074g

DO - 10.1039/b922074g

M3 - Article

C2 - 20571619

SN - 1477-0520

VL - 8

SP - 3758

EP - 3770

JO - Organic and Biomolecular Chemistry

JF - Organic and Biomolecular Chemistry

IS - 16

ER -

Isolation and characterisation of amphotericin B analogues and truncated polyketide intermediates produced by genetic engineering of Streptomyces nodosus

Abstract

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