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. 2021 May 24;26(11):3141.
doi: 10.3390/molecules26113141.

Application of Co-Culture Technology to Enhance Protease Production by Two Halophilic Bacteria, Marinirhabdus sp. and Marinobacter hydrocarbonoclasticus

Hoang Thi Hong Anh  1 , Esmaeil Shahsavari  1 , Nathan J Bott  1 , Andrew S Ball  1
Affiliations

Application of Co-Culture Technology to Enhance Protease Production by Two Halophilic Bacteria, Marinirhabdus sp. and Marinobacter hydrocarbonoclasticus

Hoang Thi Hong Anh et al. Molecules. .

Abstract

Although axenic microbial cultures form the basis of many large successful industrial biotechnologies, the production of single commercial microbial strains for use in large environmental biotechnologies such as wastewater treatment has proved less successful. This study aimed to evaluate the potential of the co-culture of two halophilic bacteria, Marinirhabdus sp. and Marinobacter hydrocarbonoclasticus for enhanced protease activity. The co-culture was significantly more productive than monoculture (1.6-2.0 times more growth), with Marinobacter hydrocarbonoclasticus being predominant (64%). In terms of protease activity, enhanced total activity (1.8-2.4 times) was observed in the co-culture. Importantly, protease activity in the co-culture was found to remain active over a much broader range of environmental conditions (temperature 25 °C to 60 °C, pH 4-12, and 10-30% salinity, respectively). This study confirms that the co-culturing of halophilic bacteria represents an economical approach as it resulted in both increased biomass and protease production, the latter which showed activity over arange of environmental conditions.

Keywords: co-culture; enzyme production; halo-bacteria; protease activity; salinity.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Cell growth (solid lines) was monitored using OD600 and extracellular protease activity (dashed lines) was determined using casein as a substrate. (a) Inoculated Marinirhabdus sp.; (b) inoculated Marinobacter sp.; (c) inoculated co-culture of Marinirhabdus sp. and Marinobacter sp. Results represent the means of three experiments, and bars indicate ± standard deviation.
Figure 2
Figure 2
Effect of temperature (a), pH (b), and salinity (c) on Marinirhabdus sp., Marinobacter sp., and coculture protease activity. Solid line—co-culture; dashed line (----)—Marinirhabdus sp.; dotted line (····)—Marinobacter sp., pH was determined at 37 °C in different buffers by varying pH values (pH 2–12; temperature was determined by assessing protease activity at different temperatures (20–60 °C); salinity was investigated in the range of 0 to 30% at 37 °C, pH 7.5. Results represent the means of three experiments, and bars indicate ± standard deviation.
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