Aims This research was conducted to investigate the biocatalytic remediation of xenobiotics polluted seawater using two biocatalysts; whole bacterial cells of facultative aerobic halotolerant Corynebacterium variabilis Sh42 and its extracted crude enzymes. Methods and Results One-Factor-at-A-Time technique and statistical analysis were applied to study the effect of initial substrate concentrations, pH, temperature, and initial biocatalyst concentrations on the batch biocatalytic degradation of three xenobiotic pollutants (2-hydroxybiphenyl (2-HBP), catechol and benzoic acid) in artificial seawater (salinity 3 center dot 1%). HPLC and gas-chromatography mass spectroscopy analyses were utilized to illustrate the quantitative removal of the studied aromatic xenobiotic pollutants and their catabolic pathway. The results revealed that the microbial and enzymatic cultures followed substrate inhibition kinetics. Yano and Koga's equation showed the best fit for the biokinetic degradation rates of 2-HBP and benzoic acid, whereas Haldane biokinetic model adequately expressed the specific biodegradation rate of catechol. The biokinetic results indicated the good efficiency and tolerance of crude enzyme for biocatalytic degradation of extremely high concentrations of aromatic pollutants than whole C. variabilis Sh42 cells. The monitored by-products indicated that the catabolic degradation pathway followed an oxidation mechanism via a site-specific monooxygenase enzyme. Benzoic acid and catechol were identified as major intermediates in the biodegradation pathway of 2-HBP, which were then biodegraded through meta-cleavage to 2-hydroxymuconic semialdehyde. With time elapsed, the semialdehyde product was further biodegraded to acetaldehyde and pyruvic acid, which would be further metabolized via the bacterial TCA cycle. Conclusion The batch enzymatic bioreactors performed superior-specific biocatalytic degradation rates for all the studied xenobiotic pollutants. Significance and Impact of the Study The enzymatic system of C. variabilis Sh42 is tolerable for toxic xenobiotics and different physicochemical environmental parameters. Thus, it can be recommended as an effective biocatalyst for biocatalytic remediation of xenobiotics polluted seawater.