Petroleum hydrocarbons are ubiquitous in nature and generally lead to contamination. The aim of this study was to establish a bio-photocatalytic system using graphite-like carbon nitride (g-C3N4) and petroleum-degrading bacterium Acinetobacter sp. JLS1 and evaluate its effect on C-16 alkane degradation. The successfully synthesized g-C3N4 produced no obvious lethal effect to strain JLS1 until 1 g/L. C-16 alkane (0.25% v/v) degradation was investigated in the individual (photocatalytic or biocatalytic) and hybrid (bio-photocatalytic) systems. The C16 alkane removal efficiency of the hybrid system under visible light irradiation was 78.2% within 4 h, which was more effective than that of photocatalytic (26.8%) and biocatalytic (38.3%) systems. This positive synergetic effect between g-C3N4 and strain JLS1 was due to the photocatalytic activity of g-C3N4 for C-16 alkane degradation and increased porosity and permeability of bacterial membrane, thus facilitating the entry of targeted compound into bacterial cells. In the hybrid system, hexadecanoic acid, dodecanoic acid and octanoic acid were examined out in bacterial cells, which were biostransformed by strain JLS1 from initial C-16 alkane and its photocatalytic degradation products (alkanes with relative lower C atoms). In addition, alkB gene transcription was strongly upregulated by g-C3N4 under visible-light irradiation, however, growth of strain JLS1 slightly improved compared to that in other tested systems, suggesting that continuous attack of photocatalytic reaction constantly impeded the repair process of bacterial cell. The established effective bio-photocatalytic system not only drew a new sight for further research but also provided a promising remediation approach for dealing with petroleum hydrocarbons. (C) 2017 Elsevier B.V. All rights reserved.