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Biosynthesis of nonribosomal peptide toxins in cyanobacteria: A functional characterisation of microcystin synthetase. Microcystins are potent toxins and tumour promoters produced by cyanobacteria associated with blue-green algal blooms. This non-ribosomal peptide is produced by microcystin synthetase, a unique enzyme complex comprised of peptide synthetases, polyketide synthases, and integrated accessory enzymes. We have identified and characterised the extensive gene cluster encoding this enzy ....Biosynthesis of nonribosomal peptide toxins in cyanobacteria: A functional characterisation of microcystin synthetase. Microcystins are potent toxins and tumour promoters produced by cyanobacteria associated with blue-green algal blooms. This non-ribosomal peptide is produced by microcystin synthetase, a unique enzyme complex comprised of peptide synthetases, polyketide synthases, and integrated accessory enzymes. We have identified and characterised the extensive gene cluster encoding this enzyme. This project describes the biochemical characterisation of specific enzyme activities within microcystin synthetase and how they determine the final structure and toxicity of the many forms of microcystin. Interactions between this enzyme complex and its substrate amino acids will provide information for the genetic engineering of this and similar natural products.Read moreRead less
BIOCATALYSTS MINED FROM CYTOCHROME P450 LIBRARIES: AN INNOVATIVE TOOL FOR ACCELERATING DRUG DEVELOPMENT. The cytochrome P450s (P450s) are a family of enzymes that are perhaps the most versatile biological catalysts known. DNA shuffling is an emerging technique that takes the genes encoding families of enzymes and creates libraries of catalysts with both improved and novel properties. We will obtain proof of concept that shuffled P450 libraries can be screened and optimized for use as biocatalys ....BIOCATALYSTS MINED FROM CYTOCHROME P450 LIBRARIES: AN INNOVATIVE TOOL FOR ACCELERATING DRUG DEVELOPMENT. The cytochrome P450s (P450s) are a family of enzymes that are perhaps the most versatile biological catalysts known. DNA shuffling is an emerging technique that takes the genes encoding families of enzymes and creates libraries of catalysts with both improved and novel properties. We will obtain proof of concept that shuffled P450 libraries can be screened and optimized for use as biocatalysts in drug development. The methodologies developed here will overcome two critical bottlenecks in current drug development: the optimisation and metabolic profiling of new drug candidates. This will yield important benefits in accelerating the optimisation and safety testing of drugs under development.Read moreRead less
Engineering of anti-platelet antibodies for the diagnosis and therapy of infants with bleeding disorders. Foeto-maternal alloimmune thrombocytopenia (FMAIT) is a serious clinical condition where infants suffer potentially fatal bleeding disorders from 14 weeks gestation to 1-2 weeks post delivery. The cause of the disease is through maternal antibodies destroying foetal platelets. Our aim is to produce human antibodies, which will be used as diagnostic agents to screen for the condition in preg ....Engineering of anti-platelet antibodies for the diagnosis and therapy of infants with bleeding disorders. Foeto-maternal alloimmune thrombocytopenia (FMAIT) is a serious clinical condition where infants suffer potentially fatal bleeding disorders from 14 weeks gestation to 1-2 weeks post delivery. The cause of the disease is through maternal antibodies destroying foetal platelets. Our aim is to produce human antibodies, which will be used as diagnostic agents to screen for the condition in pregnant women, and to further develop such antibodies for therapy. Identification of mothers at risk of FMAIT and the development of a specific therapy are vital to the management and prevention of this serious condition.Read moreRead less
The toxins of water-borne cyanobacteria: regulation and exploitation of their biosynthesis. Water quality is a major concern in Australia, as is the global need for new natural products with antibiotic activity. The mechanisms by which cyanobacteria produce toxins that reduce the quality of water may very well be the answer to the lack of novel medicinal compounds currently being discovered in nature. Encompassed in this one program are the aims of ameliorating the effects of toxic algal blooms ....The toxins of water-borne cyanobacteria: regulation and exploitation of their biosynthesis. Water quality is a major concern in Australia, as is the global need for new natural products with antibiotic activity. The mechanisms by which cyanobacteria produce toxins that reduce the quality of water may very well be the answer to the lack of novel medicinal compounds currently being discovered in nature. Encompassed in this one program are the aims of ameliorating the effects of toxic algal blooms as well as introducing the means for the design and synthesis of a range of novel bioactive products. The benefits include better water quality and biosafety management options, a new generation of drug design and discovery, and the associated transformation of environmental and medical research and education in Australia.Read moreRead less
A sustainable cellular factory for the production of antibiotics by photosynthetic bacteria. The range and rate of natural product discovery is the limiting factor in developing new pharmaceuticals. Traditional methods for the screening of these compounds or for their chemical synthesis are rapidly becoming inadequate as an increasing number of specific therapies, for cancers and infectious diseases for instance, are required. The research proposed will enable the design and production of "unnat ....A sustainable cellular factory for the production of antibiotics by photosynthetic bacteria. The range and rate of natural product discovery is the limiting factor in developing new pharmaceuticals. Traditional methods for the screening of these compounds or for their chemical synthesis are rapidly becoming inadequate as an increasing number of specific therapies, for cancers and infectious diseases for instance, are required. The research proposed will enable the design and production of "unnatural" products, including novel antibiotics, via combinatorial biosynthesis in photosynthetic microorgansims. The outcomes include graduate student training and Australian innovation in an enormous global market that is awaiting the next generation of medicines and associated pharmaceutical production technologies.Read moreRead less
Synthetic Endonucleases: Novel DNA Cleaving Agents for Cancer Chemotherapy. Cancer is a common disease in our society, with more than 1 in 4 of us dieing from it. The current survival rate is 50%, and has been so for the past 5 decades. Thus, there is clearly an urgent need for better forms of therapy. Chemotherapy is the mainstay of treatment once the disease has spread from its original site. The National Benefits from the development of a new class of effective cancer drug are two-fold. Firs ....Synthetic Endonucleases: Novel DNA Cleaving Agents for Cancer Chemotherapy. Cancer is a common disease in our society, with more than 1 in 4 of us dieing from it. The current survival rate is 50%, and has been so for the past 5 decades. Thus, there is clearly an urgent need for better forms of therapy. Chemotherapy is the mainstay of treatment once the disease has spread from its original site. The National Benefits from the development of a new class of effective cancer drug are two-fold. Firstly, much relief will come to people suffering from cancer, as some will live longer, and some will be cured. Secondly, the economic benefits are extensive, since the world market in cancer drugs is measured in billions $US, and significant monies will flow to those who hold the intellectual property rights. Read moreRead less
Enhanced biocatalysis in organic solvents for pharmaceutical biotransformation. Enzymes such as hydrolases play an important role in biotechnology because of their extreme versatility with respect to substrate specificity and stereoselectivity. The use of lipases as catalysts for optical isomer-specific organic reactions is often limited by unacceptably low enantioselectivities. We will investigate recombinant enzymes cloned from thermophilic lipolytic bacteria for synthetic reactions in orga ....Enhanced biocatalysis in organic solvents for pharmaceutical biotransformation. Enzymes such as hydrolases play an important role in biotechnology because of their extreme versatility with respect to substrate specificity and stereoselectivity. The use of lipases as catalysts for optical isomer-specific organic reactions is often limited by unacceptably low enantioselectivities. We will investigate recombinant enzymes cloned from thermophilic lipolytic bacteria for synthetic reactions in organic solvents, especially chiral resolution of mixtures in the production of pharmaceutical intermediates. Genetic improvement of lipase enantiospecificity and regioselectivity will be achieved using in vitro evolution by recombination and screening. The outcome will be cost-effective production superior biocatalysts with specifically enhanced regiospecific, enantioselective and hydrolytic characteristics.
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Bacterial monooxygenases as new biocatalysts. Using enzymes for synthesis of new pharmaceuticals is of increasing importance. Monooxygenases are particularly interesting because of their capacity to perform chemically difficult reactions thus increasing potential to make new pharmaceuticals. We have shown that an enormous pool of diverse and novel monooxygenases exists in naturally occurring bacteria. These enzymes are naturally important in pollutant degradation and regulating the emissions of ....Bacterial monooxygenases as new biocatalysts. Using enzymes for synthesis of new pharmaceuticals is of increasing importance. Monooxygenases are particularly interesting because of their capacity to perform chemically difficult reactions thus increasing potential to make new pharmaceuticals. We have shown that an enormous pool of diverse and novel monooxygenases exists in naturally occurring bacteria. These enzymes are naturally important in pollutant degradation and regulating the emissions of greenhouse gases. We will develop these enzymes for biotechnology by characterizing structural features that influence catalytic properties relevant to challenges in pharmaceutical synthesis. The outcomes will enable their engineering for specific applications in synthesis of new drugs.Read moreRead less
Venomics: Molecular and functional analysis of Australian snake venoms for development of human therapeutics. Australian snake venoms are lethal cocktails with potent effects on mammalian physiological processes, designed to immobilize and kill prey animals. Major targets of venom components are the nervous and blood coagulation systems but there is reason to believe that venoms have many other as yet unrecognized effects on mammalian systems. The project will combine techniques of modern mole ....Venomics: Molecular and functional analysis of Australian snake venoms for development of human therapeutics. Australian snake venoms are lethal cocktails with potent effects on mammalian physiological processes, designed to immobilize and kill prey animals. Major targets of venom components are the nervous and blood coagulation systems but there is reason to believe that venoms have many other as yet unrecognized effects on mammalian systems. The project will combine techniques of modern molecular biology (particularly transcriptomics and proteomics) with functional and structural analysis of purified venom components. Venoms from approximately 20 Australian snakes will be studied to reveal lead compounds for improved human pharmaceuticals against common disorders such as high blood pressure, bleeding and stroke.Read moreRead less
Directed evolution used to probe protein structure and function; new enzymes for bio-remediation and industry. The aim of the research is to generate new and useful enzymes for bio-remediation and other practical applications. For example, we are evolving enzymes to better degrade organophosphate pesticides that are environmental pollutants. Apart from producing useful enzymes, the proposed research aims at gaining a better understanding of how enzymes work and how they evolve. We intend to dete ....Directed evolution used to probe protein structure and function; new enzymes for bio-remediation and industry. The aim of the research is to generate new and useful enzymes for bio-remediation and other practical applications. For example, we are evolving enzymes to better degrade organophosphate pesticides that are environmental pollutants. Apart from producing useful enzymes, the proposed research aims at gaining a better understanding of how enzymes work and how they evolve. We intend to determine the structure of many related enzymes that have been evolved to have enhanced activities. This data will be used to analyze the intricate relationship between sequence, structure and enzyme activity.Read moreRead less