Macroalgae as feedstock for cultivation of marine bacteria

Abstract

Alginate, laminarin and mannitol amount up to 60% of dry weight in brown macroalgae. The presence of alginate and laminarin-degrading enzymes and mannitol metabolic machinery have been confirmed by Matís, a partner in European BlueGenics project. Thus, in a biorefinery perspective, R. marinus can potentially perform the saccharification and fermentation of brown macroalgae carbohydrates to yield commercial valuable biocompounds, as thermostable enzymes and glycosidic carotenoids. Rhodothermus marinus is a moderate thermophilic (65ºC) and slight halophilic (1.0% NaCl) marine bacterium. Therefore, one of the objectives of this project was to decrease the NaCl concentration in the fermentation medium, since chloride leads to a lower equipment lifetime due to stainless steel corrosion of bioreactors. The main objective of this work was the study of the bacterium R. marinus pattern of growth when cultivated in the main brown macroalgal carbohydrates. This work was performed with five R. marinus strains, two of which were successfully acclimatized to cultivation in Medium 166, cryopreserved in glycerol and recultivated in liquid media, being subject of study in the assays with different carbon and sodium sources in shake flask. The growth studies with different carbon sources suggested that (i) strain 5 presented higher glucose consumption and growth, even though none of the strains consumed all the glucose available in the media; (ii) although none of strains consumed mannitol, strain 5 seemed to be more robust to its presence; and (iii) the growth differences between the controls and the assays with alginate and pretreated alginate were not significant enough to infer if any alginate consumption occurred. It was tested a partial and total substitution of NaCl by Na2SO4. The process was not successful, since Na2SO4 seem to represent a stress factor to both R. marinus strains. Interestingly, the strain 5, when cultivated in Medium 166 containing only a half of NaCl standard concentration, presented a similar growth pattern to control. In the operational conditions imposed in shake flask cultivations containing two tested brown macroalgae (orginial and pretreated) as feedstock for growth, mannitol was not consumed. It was not possible to monitor the alginate and laminarin saccharification and fermentation. Although, the results showed that brown macroalgae are a potential feedstock under the biorefinery concept, since some R. marinus growth was observed. The more promising result to BlueGenics project was obtained from shake flask cultivations of strain 5 in Medium 166 with 0.500% NaCl and 10.0 g.L-1 glucose, since the growth with low chloride content determinates the feasibility of the scale-up of the process to bioreactor . Because of that, the assay was validated in 3L controlled bioreactor. The process presented a μmax of 0.208 h-1, a maximum biomass concentration of 8.75 gX.L-1, a volumetric biomass production rate of 0.295 g.L-1.h-1 and a volumetric glucose uptake rate of 0.293 g.L-1.h-1. Some feeding strategies were tested but further assays have to be performed in order to optimize the bioprocess.