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The present study investigates the conversion efficacy of glucose into calcium salt of gluconic acid (calcium gluconate) through fermentation by using the fungus Aspergillus niger (A. niger). The study further evaluates the impact of various process parameters and thereby identifies the optimum parameters to maximize the output. The different process parameters evaluated in this study were glucose concentration, pH, temperature, and aeration. After that, the optimized process pre-treatment parameters were used to determine the feasibility of the process for further industrial scale-up. The obtained results indicated enough competency of the optimized parameters to be reproduced at the industrial level. The obtained product was expected to meet the requirement as per the specifications led by various industrial requirements like food and feed, pharmaceutical industry, aquaculture and food industries, etc. The overall scheme of work presented in this study will help to define the techno-economic viability (in terms of energy consumption, environmental concerns, and higher yield) of the process and thereby will benefit various stakeholders.