Awards/ Honors:

• Award of PhD scholarship under overseas scholarship program by Higher Education Commission of Pakistan.
• Award of Post Doctoral fellowship by DSM anti-infective and GBB under STW program, The Netherlands (2012-2014).
• Member of Klyver Center for Genomic and Proteomics, Delft University of Science and Technology, The Netherlands.

Academic Qualifications:

Research Interests

1- Industrial scale production of anti-fungal compounds

Antifungals have an effective role in protecting humans from pathogenic microorganisms. Iturin A is the cyclic lipopeptide, produced by Bacillus spp, has demonstrated broad spectrum antifungal and anticancer activities. However, its low yield, high production cost, and toxicity are the major issues in pursuing the discovery/development of this lipopeptide as the antifungals / anticancer agents at industrial scale. We aim to genetically engineer the biosynthesis pathway of Iturin A by rearranging its domain/modules to produce analogues with unique structural and biological properties.

Iturin A is a cyclic lipopeptide, produced by Bacillus spp. Structurally, Iturin A consists of two parts, a cyclic peptide (L-Asn–D-Tyr–D-Asn–L-Gln–L-Pro– D–Asn–L-Ser) and hydrophobic tail of fatty acid composition. The cyclic peptide is biosynthesized via Non ribosomal peptide synthetase mechanism (ituA, ituB and ituC). Iturin-A has wide-scale and broad spectrum antifungal activity and quite recently this lipopeptide also demonstrated anticancer activity. Hence, can be used as an effective remedy for both human and animal pathogenic mycoses as well as against other ailments such as cancer.

The proposed project aims to develop methodologies for enhance production of iturin A analogues with reduced toxicity after tuning the biosynthesis pathway. Here we plan to exploit the superb fermentation characteristics of B. subtilus in conjunction with enzyme engineering approaches for the production of a variety of natural and unnatural iturin-A related compounds. We will generate a host strain that lacks the entire fengycin and surfactin gene clusters, and re-introduce the complete and incomplete iturin-A biosynthetic gene on a plasmid to enable high level production of various intermediates and end products.

In addition, we will rearrange the adenylation domain schematically for the biosynthesis of modified final products and intermediates with the aim to improve upon the antimicrobial activity and to reduce toxicity¹⁰. Advanced analytics (LC-MS/MS) will be employed to characterize the purified compounds that will be thoroughly tested for antimicrobial screening.

2- Neutraceuticals:

In Pakistan Antibiotics are used as Growth Promoters (AGP) particularly in poultry. Exploitation of these antibiotics as growth promoter resulted in Multidrug Resistant Super Bugs (MRSB). European Union introduced a total ban on antibiotics as growth promoter in 2006. Due to the use of AGP in poultry, Pakistan cannot export poultry meat. Similarly, Pakistan poultry products cannot reach to the Halal food market which has total worth of $3 trillion in the world (Economic survey, 2018). Halal meat contributes about 16 percent of total world trade of the commodity.

Industrial Enzymes and Biofuel group at National Institute of Biotechnology and Genetics Engineering, Faisalabad has successfully developed a microbial consortium that showed remarkable results in poultry as an alternative to Antibiotics and as Growth Promoter (support letter from Industry can be available). This microbial consortium consists of microorganism like Pichia pastoris, Saccharomyces cerevisiae, Aspergillus oryzae and Actinomycetes species that are GRAS. Our preliminarily analysis has indicated that by introducing this consortium and its variants in poultry industry, Antibiotics as growth promoters can be replaced, thus allowing our poultry products in Halal as well in international market. Currently, Poultry sector in Pakistan is a source of income for about 2.5 million people and the contribution of poultry meat to the total meat production of Pakistan is 28% (Pakistan’s Economic Survey-2018). Our poultry market can attain export acquiescence only if we can switch to antibiotics free production using nutraceuticals including Direct Feed Microorganism (DFM), feed additives, growth promoters and the drugs that are herbal in nature.

Selected Publications:

1. Bushra Aleem, M. Hamid Rashid, Neelam Zeb, Anum Saqib, Aisha Ihsan, Mazhar Iqbal, Hazrat Ali. (2018) Random mutagenesis of super Koji (Aspergillus oryzae): improvement in production and thermal stability of α-amylases for maltose syrup production. BMC Microbiol. 2018 Nov 28;18(1):200. doi: 10.1186/s12866-018-1345-y
2. Viggiano A, Salo O, Hazrat Ali, Szymanski W, Lankhorst PP, Nygård Y, Bovenberg RAL4,5, Driessen AJM. (2017) Elucidation of the biosynthetic pathway for the production of the pigment chrysogine by Penicillium chrysogenum. Appl Environ Microbiol. doi: 10.1128/AEM.02246-17
3. Hazrat Ali, Marco Ries, Jeroen.G. Nijland, Peter Lankhorst, Thomas Hankemeier, Roel A.L. Bovenberg, Rob J. Vreeken, Arnold J.M. Driessen^. (2015) Novel key metabolites reveal further branching of the roquefortine/meleagrin biosynthetic pathway. Journal of Biological chemistry doi: 10.1074/jbc.M113.512665
4. Hazrat Ali, Marco Ries, Jeroen.G. Nijland, Peter Lankhorst, Thomas Hankemeier, Roel A.L. Bovenberg, Rob J. Vreeken, Arnold J.M. Driessen^. (2014) A non-canonical NRPS is involved in the synthesis of fungisporin and related hydrophobic cyclic tetrapeptides in Penicillium chrysogenum. PLoS One. 2014 Jun 2;9(6):e98212. doi: 10.1371/journal.pone.0098212
5. Hazrat Ali, Marco Ries, Jeroen.G. Nijland, Peter Lankhorst, Thomas Hankemeier, Roel A.L. Bovenberg, Rob J. Vreeken, Arnold J.M. Driessen^. (2014) A branched biosynthetic pathway is involved in production of roquefortine and related compounds in Penicillium chrysogenum. PLoS ONE 8(6): e65328. doi:10.1371/journal.pone.0065328