Part B | 14.3
418 Part B Tools and Methods in Marine Biotechnology
toga maririma [14.34]. The putative enzyme had no
amino acid sequence similarity to the glycoside hydrolase family (GHF) 13 ˛-amylases, yet the range of
substrate hydrolysis and the product profile suggest the
protein to be an ˛-amylase. A marine Bacillus was reported to produce several ˛-amylases that can degrade
various types of plant-based raw starch to produce simple sugars [14.35, 36]. Two statistical analyses were
applied to optimize the media composition and fermentation parameters for cold adapted amylase production
by the marine isolate Wangia sp. C52 in a shake flask
culture [14.33]. The study showed starch, tryptone, and
initial pH had significant effects on production of this
enzyme. The maximum amylase production was optimized at 20
ı C and pH 7:18 with a shaking speed
of 180 rpm. The experimental yield of the amylase in
the optimized media closely matched the yield predicted by the statistical model when the fermentation
was performed at theoretically optimized fermentations conditions. Tenfold higher amylase production
was observed after this optimization. Chakraborty et al.
isolated two thermostable alkaliphilic amylases from
marine microorganisms that showed excellent stability
towards surfactants and commercial detergents – the
enzymes, therefore, have potential applications in the
detergent industry [14.37, 38].
Cellulases and Lignocellulases
Cellulolytic enzymes were reported from a range of
marine microorganisms, including bacteria and fungi.
A Saccharophagus degradans strain was reported
to degrade large numbers of complex polysaccharides from numbers of algae, plant, and invertebrate
sources [14.39]. Complete genome sequencing of the
bacteria revealed more than 180 open reading frames
encoding carbohydrate degrading enzymes. It appears
that this bacterium consists of a full array of polysaccharide degrading enzymes for complete degradation
of plant cell walls. 13 cellulose depolymerases and
seven accessory enzymes including two cellodextrinases, three cellobiases, a cellodextrin phosphorylase,
and a cellobiose phosphorylase were predicted by genomic and proteomic analyses. A solvent stable alkaline
cellulase was reported from a solvent tolerant marine
strain of Bacillus aquimaris [14.40]. Enzyme stability increased with the presence of 20 v=v % benzene
and it was significantly stable in the same concentration of methanol, acetone, and toluene. The enzyme
activity was increased to 150% and 155%, respectively, by pre-incubation in two ionic liquids 1-ethyl3-methylimidazolium methanesulfonate and 1-ethyl-3methylimidazolium bromide. Cellulase productions by
a group of marine microorganisms were optimized using industrial and agricultural wastes. Rice bran was reported to be a preferential nutrient for optimum production of three cellulases by a Psychrobacter aquimaris
strain [14.41]. The microbe has optimum growth at
25
ı C but maximum production of avicelase, CMCase,
and filter paperase was achieved at 30
ı C. Aeration was
more important than agitation for cell growth and fermentation of all three cellulases. Enzyme production
was scaled up to a 100 l pilot plant bioreactor. El-Sersy
et al. screened six marine strains of actinomycetes for
CMCase production and a Streptomyces ruber strain
showed the best productivity [14.42]. Rice straw was
used as the carbon source. The Plackett–Burman design was applied to optimize the media composition
and fermentation parameters for maximum enzyme
production. Two different cellulases were partially purified and their specific activities were determined as
4239:697 U mg
1 and 846:752 U mg
1 , respectively.
Three alkaline cellulase producing fungal strains were
isolated from mangrove leaves and wood litters [14.43].
The production of ˇ-endoglucanase, ˇ-exoglucanase,
and ˇ-glucosidase by these isolates was optimized in
submerged fermentation and solid-state fermentation
using cheap agricultural and industrial waste as substrates. Maximum enzyme production was achieved by
solid-state fermentation at alkaline pH using media containing cotton seed. It is interesting to note that the
enzymes produced under alkaline conditions showed
higher stability and activity than those produced by the
same organisms under non-alkaline conditions. Pointing et al. studied 15 marine fungi for cellulolytic and
lignolytic activity [14.44]. The lignolytic activity of
these microorganisms was dependent on the primary
carbon source, pH, and salinity of the medium. In
another study, 29 marine fungi were screened for lignocellulose-degrading enzyme activities in agar media [14.45]. The isolates were identified as 27 ascomycetes, one basidiomycete (Calathella mangrovei),
and a mitosporic fungus (Cirrenalia tropicalis). Cellulolytic, xylanolytic, and ligninolytic activities were
studied in all 29 isolates. 21 isolates produced three enzymes: endoglucanase, xylanase, and laccase, while 2
isolates showed a lack of xylanase activity and 6 isolates had no laccase activity.
Chitinases
Chitin is a major component of fungal cell walls, the
exoskeleton of arthropods including insects and crustaceans (mollusks, crabs, lobsters, and shrimps), and the
418 Part B Tools and Methods in Marine Biotechnology
toga maririma [14.34]. The putative enzyme had no
amino acid sequence similarity to the glycoside hydrolase family (GHF) 13 ˛-amylases, yet the range of
substrate hydrolysis and the product profile suggest the
protein to be an ˛-amylase. A marine Bacillus was reported to produce several ˛-amylases that can degrade
various types of plant-based raw starch to produce simple sugars [14.35, 36]. Two statistical analyses were
applied to optimize the media composition and fermentation parameters for cold adapted amylase production
by the marine isolate Wangia sp. C52 in a shake flask
culture [14.33]. The study showed starch, tryptone, and
initial pH had significant effects on production of this
enzyme. The maximum amylase production was optimized at 20
ı C and pH 7:18 with a shaking speed
of 180 rpm. The experimental yield of the amylase in
the optimized media closely matched the yield predicted by the statistical model when the fermentation
was performed at theoretically optimized fermentations conditions. Tenfold higher amylase production
was observed after this optimization. Chakraborty et al.
isolated two thermostable alkaliphilic amylases from
marine microorganisms that showed excellent stability
towards surfactants and commercial detergents – the
enzymes, therefore, have potential applications in the
detergent industry [14.37, 38].
Cellulases and Lignocellulases
Cellulolytic enzymes were reported from a range of
marine microorganisms, including bacteria and fungi.
A Saccharophagus degradans strain was reported
to degrade large numbers of complex polysaccharides from numbers of algae, plant, and invertebrate
sources [14.39]. Complete genome sequencing of the
bacteria revealed more than 180 open reading frames
encoding carbohydrate degrading enzymes. It appears
that this bacterium consists of a full array of polysaccharide degrading enzymes for complete degradation
of plant cell walls. 13 cellulose depolymerases and
seven accessory enzymes including two cellodextrinases, three cellobiases, a cellodextrin phosphorylase,
and a cellobiose phosphorylase were predicted by genomic and proteomic analyses. A solvent stable alkaline
cellulase was reported from a solvent tolerant marine
strain of Bacillus aquimaris [14.40]. Enzyme stability increased with the presence of 20 v=v % benzene
and it was significantly stable in the same concentration of methanol, acetone, and toluene. The enzyme
activity was increased to 150% and 155%, respectively, by pre-incubation in two ionic liquids 1-ethyl3-methylimidazolium methanesulfonate and 1-ethyl-3methylimidazolium bromide. Cellulase productions by
a group of marine microorganisms were optimized using industrial and agricultural wastes. Rice bran was reported to be a preferential nutrient for optimum production of three cellulases by a Psychrobacter aquimaris
strain [14.41]. The microbe has optimum growth at
25
ı C but maximum production of avicelase, CMCase,
and filter paperase was achieved at 30
ı C. Aeration was
more important than agitation for cell growth and fermentation of all three cellulases. Enzyme production
was scaled up to a 100 l pilot plant bioreactor. El-Sersy
et al. screened six marine strains of actinomycetes for
CMCase production and a Streptomyces ruber strain
showed the best productivity [14.42]. Rice straw was
used as the carbon source. The Plackett–Burman design was applied to optimize the media composition
and fermentation parameters for maximum enzyme
production. Two different cellulases were partially purified and their specific activities were determined as
4239:697 U mg
1 and 846:752 U mg
1 , respectively.
Three alkaline cellulase producing fungal strains were
isolated from mangrove leaves and wood litters [14.43].
The production of ˇ-endoglucanase, ˇ-exoglucanase,
and ˇ-glucosidase by these isolates was optimized in
submerged fermentation and solid-state fermentation
using cheap agricultural and industrial waste as substrates. Maximum enzyme production was achieved by
solid-state fermentation at alkaline pH using media containing cotton seed. It is interesting to note that the
enzymes produced under alkaline conditions showed
higher stability and activity than those produced by the
same organisms under non-alkaline conditions. Pointing et al. studied 15 marine fungi for cellulolytic and
lignolytic activity [14.44]. The lignolytic activity of
these microorganisms was dependent on the primary
carbon source, pH, and salinity of the medium. In
another study, 29 marine fungi were screened for lignocellulose-degrading enzyme activities in agar media [14.45]. The isolates were identified as 27 ascomycetes, one basidiomycete (Calathella mangrovei),
and a mitosporic fungus (Cirrenalia tropicalis). Cellulolytic, xylanolytic, and ligninolytic activities were
studied in all 29 isolates. 21 isolates produced three enzymes: endoglucanase, xylanase, and laccase, while 2
isolates showed a lack of xylanase activity and 6 isolates had no laccase activity.
Chitinases
Chitin is a major component of fungal cell walls, the
exoskeleton of arthropods including insects and crustaceans (mollusks, crabs, lobsters, and shrimps), and the