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18 Advancements in Bio-hydrogen Production from Waste Biomass
by photo-fermentation [35]. Although the literature includes a large number of
studies on the batch mode of H2 production, but it has some disadvantages over
other modes of operations. The culture conditions are highly unstable due to the
periodic removal of the sample. Moreover, due to the closed system, the removal of
extracellular products is not possible, which results in product inhibition by liquid
and gaseous metabolites.
18.4.5.2
Continuous Mode
The continuous mode of operation is often favored for biohydrogen production
from various strains. Chemostat is characterized by a constant supply of cul-
ture medium or substrate to the reactor, which maintains the microbes in the
exponential growth phase for prolonged periods and regular removal of product
streams. A continuous stirred tank reactor (CSTR) is widely used for both photo
and dark fermentation. Rhodopseudomonas and Clostridium are cultivated in CSTR
for photo- and dark-fermentation, respectively [36]. Kim and Lee (2010) have
used a microbial consortium in CSTR to obtain 2.3 mol-H2/mol-glucose, whereas
3.4 mol-H2/mol-glucose was obtained by using anaerobic sludge [37, 38]. The
chemostat mode of fermentation is also used to enhance the yield of biohydrogen
from 5.8 to 11.61 mol-H2/mol-sucrose by varying the pH of the media and cultiva-
tion temperature. To further increase the biohydrogen production using CSTR, the
use of biofilms or immobilized organisms has been reported previously [39].
18.4.5.3
Fed Batch
The fed-batch mode of fermentation involves the periodic addition of substrate
to the reactor, followed by the removal of the product only after completion of a
reaction cycle. This mode of fermentation ensures that there is no saturation or
substrate inhibition. The literature survey suggested that only a few reports on
fed-batch dark fermentative hydrogen production as compared to other modes of H2
production. This is mainly due to the massive formation of organic acids, alcohols,
and other metabolites in the reactor, which lowers the kinetics of the process and
also inhibits the growth of the microorganism. The fed-batch fermentation has
reported up to 3.1 mol-H2/mol-glucose using microbial consortium and a yield of
2.15 mol-H2/mol-substrate using a recombinant strain of Clostridium [32]. The
potential of the fed-batch process for efficient production of fermentative H2 needs
further detailed investigations in terms of quantity and quality.
18.5
Strategies to Enhance Microbial Hydrogen
Production
The earlier studies on the improvement of biohydrogen production have suggested
several approaches, such as statistical optimization of process parameters and
fermentation medium, reconstruction of a metabolic network by metabolic flux
analysis (MFA), genetic engineering of microorganisms, and improved fermenta-
tion kinetics by application of ultrasound. This section brings forward the strategies
in detail and describes their potential to enhance microbial H2 production.