Water being vital for all life forms is an utmost important molecule. Industrial and pharmaceutical waste holds 80% of the total water pollutants of the world. Among various water remediation techniques biosorption has its own effective solution, it being economic, zero waste emission, and no heavy machinery requirement. Binary and multi-mode biosorption studies are used to map the interaction pathway. The phenomenological mathematical modeling of the kinetics of biosorption that will be used in this study is based on equilibrium equations, material balances and possible limiting mass transfer steps instead of the empirical models usually applied. The model is then adjusted to the experimental kinetic data and validated by a simulation in a different condition or sample model. Regeneration study determines the marketability of the adsorbent on large scale. Interaction pathway to verify how adsorbate and adsorbent behave in their micro-environment is one of such often neglected areas in biosorption and it is quite challenging. We will be using mathematical modeling to elaborately study these points. Apart from using a simulated water model, real effluent will be collected and validated for the nano-biosorbent for its versatility in both batch as well as continuous column studies.

In accordance with PAT initiative, the CPPs/CQAs of the process need to be identified and monitored in real-time mode to achieve improved product quality (FDA, 2004). The CPPs governing glycan moiety and protein productivity was already reported in several articles. But deconvolution of soft sensor’s/online sensor’s input into CPPs are at the nascent stage for most of the therapeutic protein production. Moreover, several therapeutic protein products in the Indian biopharma sector are under pipeline to develop suitable PAT platform for an effective monitoring and control. Incorporation of multiple soft sensors to assess different process parameters and ascertain various stages of growth and production formation would provide the glimpses of the carbon substrate channelled towards various metabolic pathways. Cultivation of organism carried out by manipulating μ was determined to have stronger influence at their metabolic level. P. pastoris when operated at retentostat i.e. operating the organism at nearly zero specific growth rate (μ < 0.001 h-1), it leads to the transcriptional re-programming followed by the up-regulation of stress related genes and down-regulation of cell cycle machinery. For all growth associated products, it is well established about the proportionality constant appeared to be conserved for a typical μ & 𝑞𝑃 values. This ratio represents the stoichiometric coefficient of the sequence of all pipe-line events in the protein production machinery. Therefore, aiming the control of μ (critical process parameter) at a process level will lead to a more promising product output.

Heparosan is most commonly known as a precursor for the widely used anticoagulant, heparin. It is present and procured from mainly bovine lung tissue, sheep lung tissue and porcine intestine tissue. Some novel properties of heparosan such as moisture retention capacity, general non-immunogenicity and biocompatibility have allowed it to be used in a multitude of applications, such as being a dermal filler, drug delivery applications, the ability to be used as a biomaterial, ophthalmologic capabilities, and as medical device coatings. Many efforts have been made to synthesize heparin to overcome the side effects and insufficient supply of heparin. In this study, a GRAS organism, Lactococcus Lactis, is used as a host organism with heparosan producing genes from E. coli K5. Lactococcus Lactis has shown to be a good host with a GRAS status and with many well-defined genetic tools available. One of the most popular expression systems in L. lactis involves the employment of a nisin-inducible promoter in a nisin-negative strain such as L. lactis NZ9000, which has the nisRK genes integrated into its chromosome. The nisin-controlled expression (NICE) system is a very tightly controlled, strong expression system which shows a linear dose– response between inducer concentration at subinhibitory level and protein production, which is used in this study for the expression of the genes of interest. The genes of interest used are KfiA and KfiC, which have been taken from E.coli K5 genomic DNA and have glycosyltransferase activities which determined to be essential for heparosan synthesis, and   ugd, glmU which is responsible for the synthesis of UDP-glucose dehydrogenase and UDP-N-acetylglucosamine. Studies to determine the effect of addition of compounds like glutamate and acetate to enhance precursor concentration its effect on final product concentration which will be carried out in batch and fed batch reactor runs, as well as optimizing media and purification and characterization of final product and precursor concentrations using 1-D NMR spectroscopy.

Wax ester (WE) is a type of high-value neutral lipid formed by the conjugation of fatty acid and fatty alcohol.  It has numerous applications such as high-grade lubricants, wound healing, bio-surfactants, gum-bases, personal care products, biodiesel production, etc. Depending on the carbon length and the degree of unsaturation, its property differs (oxidation stability, melting point) and hence the end application is disparate. The predominant animal and plant waxes are beeswax, spermaceti oil and jojoba (Simmondsia chinensis) oil respectively. The global ban on whale hunting and the high cost of jojoba oil production were being the stumbling blocks for its commercialization. A few decades back industries opted for chemical synthesis but it suffered the disadvantage of using corrosive acids, hazardous chemical handling, synthetic ester degradation and energy requirement was also high. A promising and possible replacement for chemical synthesis is the use of the biotechnological approach (immobilized lipase esterification). Though the use of a solvent-free immobilised system was accentuated because of its high conversion yield, a major burden that hinder us to say it an efficient process is the external supply of synthetic fatty alcohol and fatty acid.  Algae is an efficient lipid producer but its growth rate and productivity are 25 folds and 75-folds lower than bacteria. Hence we have chosen a microbial platform for its production. . In silico metabolic engineering of the host organism to identify the genes that would increase the flux towards WE production rather than its competing pathways is in progress. Bioprocess optimization studies are underway utilizing recombinant WE producing strain to achieve high productivity.