Penicillium Metabolic Engineering Services

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Penicillium metabolic engineering services offer specialized solutions for optimizing the metabolic pathways of Penicillium species to enhance the production of valuable biochemicals, pharmaceuticals, enzymes, and other industrially relevant products. Our comprehensive services provide support from initial project design to final strain optimization, ensuring precise and efficient metabolic modifications tailored to your specific research and biotechnological needs.

Integration of the calbistrin cluster from P. decumbens into 4xKO-B and verifcation of production. (C Pohl, et al.,2020)

Overview Service Process Examples and Solutions Case Study Frequently Asked Questions

Overview

Penicillium species, including Penicillium chrysogenum and Penicillium roqueforti, are widely used in biotechnology for their ability to produce antibiotics, enzymes, and secondary metabolites. Our metabolic engineering services leverage advanced genetic engineering techniques, such as CRISPR/Cas9, homologous recombination, and synthetic biology approaches, to optimize the metabolic pathways of Penicillium for improved production of target compounds.

Service Process

The process of Penicillium metabolic engineering involves several critical and interrelated steps:

Project Consultation: Collaborating with researchers to define specific metabolic engineering goals, including target compounds, desired metabolic modifications, and intended applications.
Pathway Analysis and Design: Analyzing existing metabolic pathways and designing modifications to optimize the production of target compounds. This includes pathway reconstruction and flux balance analysis.
Vector Design and Construction: Designing and constructing expression vectors or CRISPR/Cas9 systems tailored to the specific genetic modifications needed for the metabolic pathway.
Fungal Transformation: Introducing the genetic material into Penicillium cells using techniques such as protoplast transformation, Agrobacterium-mediated transformation, or electroporation.
Selection and Screening: Selecting successfully transformed cells using selectable markers and screening for desired metabolic modifications using assays such as HPLC, GC-MS, and enzymatic assays.
Strain Optimization: Optimizing the engineered strains through iterative rounds of modification and selection to enhance the production of target compounds. This may include optimizing growth conditions and media composition.
Characterization and Validation: Characterizing the engineered strains to confirm the presence and functionality of the metabolic modifications. This includes growth assays, metabolic profiling, and functional assays.
Scale-Up and Production: Scaling up the engineered strains for large-scale production and further applications in research or industry.
Reporting and Consultation: Providing a detailed report of the findings and offering further consultation to interpret the results and plan subsequent research steps.

For more information about our Penicillium Metabolic Engineering Services or to discuss your specific needs, please contact us. Our team of experts is available to provide guidance and support for your research and biotechnological projects, ensuring you achieve your scientific and industrial goals.

Examples and Solutions

The following table provides an overview of various case studies in Penicillium metabolic engineering and the solutions we offer to support your research and biotechnological endeavors:

Case Study	Description	Solutions We Offer
Antibiotic Production Enhancement	Engineering Penicillium strains to increase the yield of antibiotics such as penicillin.	Pathway design, genetic modification, strain optimization, and scale-up.
Statin Production Optimization	Modifying metabolic pathways to enhance the production of statins.	CRISPR/Cas9 gene editing, pathway optimization, and production scaling.
Organic Acid Production	Developing strains for the efficient production of organic acids for industrial use.	Metabolic pathway reconstruction, strain engineering, and yield optimization.
Industrial Enzyme Production	Engineering Penicillium to produce high levels of industrial enzymes.	Pathway design, strain development, and production optimization.
Biochemical Synthesis	Engineering Penicillium to produce solvents and other industrial biochemicals.	Synthetic biology, pathway integration, and functional assays.
Synthetic Pathway Construction	Constructing synthetic pathways in Penicillium for the production of novel biochemicals.	Synthetic biology, pathway integration, and functional assays.

Case Study

Penicillium chrysogenum was successfully engineered to produce a novel carbamoylated cephalosporin that can be used as a synthon for semi-synthetic cephalosporins. To this end, genes for Acremonium chrysogenum expandase/hydroxylase and Streptomyces clavuligerus carbamoyltransferase were expressed in a penicillinG high-producing strain of P. chrysogenum. Growth of the engineered strain in the presence of adipic acid resulted in production of adipoyl-7-amino-3-carbamoyloxymethyl3-cephem-4-carboxylic acid (ad7-ACCCA) and of several adipoylated pathway intermediates. A combinatorial chemostat-based transcriptome study, in which the ad7-ACCCA-producing strain and a strain lacking key genes in b-lactam synthesis were grown in the presence and absence of adipic acid, enabled the dissection of transcriptional responses to adipic acid per se and to ad7-ACCCA production. Transcriptome analysis revealed that adipate catabolism in P. chrysogenum occurs via b-oxidation and enabled the identification of putative genes for enzymes involved in mitochondrial and peroxisomal b-oxidation pathways. Several of the genes that showed a specifically altered transcript level in ad7- ACCCA-producing cultures were previously implicated in oxidative stress responses

ad7-ACCCA production in shake flasks(DM Harris, et al.,2009)

ad7-ACCCA production in shake flasks. (A) NMR spectrum of ad7ACCCA-producing DS17690 P. chrysogenum transformants. (B) Relative stability of ad7-ACCCA at various conditions. The pH of ad7-ACCCA-containing filtrates was adjusted with 4 N HCl or 4 N KOH to either pH 4 or 13. The samples were then incubated for 8 h at 4, 25 and 80 1C and subsequently analysed by NMR. (C) Relative concentration distribution of ad7ADACCCA and intermediates in shake-flask experiments after 7 days of cultures. The experiment was performed in triplicate and the standard error did not exceed 10%.

Frequently Asked Questions

Q: What is Penicillium metabolic engineering?

A: Penicillium metabolic engineering involves the genetic modification of Penicillium strains to optimize their metabolic pathways for the production of target compounds. This can include introducing, deleting, or modifying specific genes to redirect metabolic fluxes and increase the yield of desired products.

Q: How is Penicillium metabolic engineering performed?

A: Penicillium metabolic engineering is performed through a series of steps including project consultation, pathway analysis and design, vector design and construction, fungal transformation, selection and screening, strain optimization, characterization and validation, scale-up and production, and reporting. Each step ensures precise and efficient metabolic modifications.

Q: What are the applications of Penicillium metabolic engineering?

A: Applications include antibiotic production, pharmaceutical production, industrial enzyme production, biochemicals production, agricultural biotechnology, and synthetic biology. Engineered Penicillium strains are used to produce valuable bioproducts and address various industrial and environmental challenges.

Q: What are the key steps in the Penicillium metabolic engineering process?

A: Key steps include project consultation, pathway analysis and design, vector design and construction, fungal transformation, selection and screening, strain optimization, characterization and validation, scale-up and production, and reporting. These steps ensure comprehensive and accurate development of engineered Penicillium strains.

Q: Why is Penicillium metabolic engineering important?

A: Penicillium metabolic engineering is important for advancing research, developing new bioproducts, optimizing industrial processes, and addressing environmental challenges. Engineered Penicillium strains provide valuable tools for enhancing production yields and creating novel compounds.

Please note that all services are for research use only. Not intended for any clinical use.

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