Isolation, Screening, and Evaluation
of Cellulase-Producing Bacteria
from the Soil of Similipal Biosphere
Reserve for Biofuel Production
from Lignocellulosic Biomass
Manish Paul, Soumya Ranjan Meher, Subhadarshini Giri,
and Hrudayanath Thatoi
Abstract Lignocellulose is found in nature as an alternative source of energy and
can be used for the production of bioethanol. Cellulose, one of the most abundant
components of lignocellulose can be hydrolyzed using enzymes cellulase to produce
glucose, which can be used for the production of ethanol. Cellulase production from
cellulolytic bacteria is challenging. Hence, research has been focused on isolation and
identification of efficient cellulolytic bacteria for their use in bioethanol production.
In the present investigation, the most potent screened bacterial strain was subjected
to optimization of its cellulase production by using response surface methodology
taking four independent variables such as substrate concentration, pH, temperature,
and incubation time. The optimization result showed that the bacteria had a maximum
production of cellulase enzyme of 617.71 U/mL in an optimized condition at an
incubation time of 42 h, pH 9, carboxy methyl cellulose (CMC) concentration of 15
gm/L, and temperature of 37.5 °C. The strain, SCB9, was identified as Bacillus albus
based on the 16S rRNA sequencing and phylogeny analysis. Overall results from this
study indicate that the cellulolytic bacteria SCB9 is a potent candidate for cellulase
production, which can be exploited for bioethanol production from lignocellulosic
biomass through adaptation of further appropriate biotechnological approaches.
Keywords Lignocellulosic biomass · Similipal biosphere reserve · Cellulolytic
bacteria · Cellulase · Response surface methodology · Enzyme optimization
M. Paul · S. R. Meher · S. Giri · H. Thatoi (B)
Department of Biotechnology, North Orissa University, Takatpur, Baripada 757003, Odisha, India
e-mail: hn_thatoi@rediffmail.com
M. Paul
e-mail: p.manish88@gmail.com
S. R. Meher
e-mail: soumyameher222@gmail.com
S. Giri
e-mail: www.shubhadarshini@gmail.com
© Springer Nature Singapore Pte Ltd. 2021
D. Ramkrishna et al. (eds.), Advances in Bioprocess Engineering and Technology,
Lecture Notes in Bioengineering,
https://doi.org/10.1007/978-981-15-7409-2_46
457
of Cellulase-Producing Bacteria
from the Soil of Similipal Biosphere
Reserve for Biofuel Production
from Lignocellulosic Biomass
Manish Paul, Soumya Ranjan Meher, Subhadarshini Giri,
and Hrudayanath Thatoi
Abstract Lignocellulose is found in nature as an alternative source of energy and
can be used for the production of bioethanol. Cellulose, one of the most abundant
components of lignocellulose can be hydrolyzed using enzymes cellulase to produce
glucose, which can be used for the production of ethanol. Cellulase production from
cellulolytic bacteria is challenging. Hence, research has been focused on isolation and
identification of efficient cellulolytic bacteria for their use in bioethanol production.
In the present investigation, the most potent screened bacterial strain was subjected
to optimization of its cellulase production by using response surface methodology
taking four independent variables such as substrate concentration, pH, temperature,
and incubation time. The optimization result showed that the bacteria had a maximum
production of cellulase enzyme of 617.71 U/mL in an optimized condition at an
incubation time of 42 h, pH 9, carboxy methyl cellulose (CMC) concentration of 15
gm/L, and temperature of 37.5 °C. The strain, SCB9, was identified as Bacillus albus
based on the 16S rRNA sequencing and phylogeny analysis. Overall results from this
study indicate that the cellulolytic bacteria SCB9 is a potent candidate for cellulase
production, which can be exploited for bioethanol production from lignocellulosic
biomass through adaptation of further appropriate biotechnological approaches.
Keywords Lignocellulosic biomass · Similipal biosphere reserve · Cellulolytic
bacteria · Cellulase · Response surface methodology · Enzyme optimization
M. Paul · S. R. Meher · S. Giri · H. Thatoi (B)
Department of Biotechnology, North Orissa University, Takatpur, Baripada 757003, Odisha, India
e-mail: hn_thatoi@rediffmail.com
M. Paul
e-mail: p.manish88@gmail.com
S. R. Meher
e-mail: soumyameher222@gmail.com
S. Giri
e-mail: www.shubhadarshini@gmail.com
© Springer Nature Singapore Pte Ltd. 2021
D. Ramkrishna et al. (eds.), Advances in Bioprocess Engineering and Technology,
Lecture Notes in Bioengineering,
https://doi.org/10.1007/978-981-15-7409-2_46
457