The Process of Xylanase Production from Bacillus pumilus RXAIII-5
Volume I, Numbti 2
The Process of Xylanase Production from
Baci~mpumilusRXAIII-5
NUR RICHANA1$*,TUN TEDJA IRAWADI', M. ANWAR NUR',
ILLAH SAILAH2, m KHASWAR SYAMSU2
IDepartment of Chemistry, F a d @ of Ydhemafics and Naiuml S d e n m
'DqmfmentofAgrolnhstdal TechnoIogy, F~acwflyof Agrolmrlrrsfrial Technology
Insiitri Pemnian Bogor, Darmaga Cartrprrs, Bogor 16680,Indonesiu
The optlmum conditions for the growth or Bsclllns pnmllus R X A I I I - 5 (a potential xylanist producer) were
nought. tbtst included trmpcraturc, pH, aeration. and agitation of the calturt batcb. Afterwards a mathematical
modtl bawd on the parameter of cultivation klnttics was formulattd. At the uamc time, the rheology o f the fluid
a d COT battcrlal crltlvatlon in a biortactor was studied. The data obtained was used for cstlmatlng the 'scaling
tip' of tnzyme production. The results of tbc study indicate thmt the optimum rondltion for proceraing I n 50 ml
E r l t n m t y t r flask a r t used temperature of 35 'C (308'K), pH 7, and an agitatlon rate or 140 rpm. The bigbest
xylama8c activity aod Its sprciflr activity arc 297.132 U,ml-I mod 655.32 U.g-'profitin, rtnptctlvtly. Subsequent
cxperimtntn In a blorcactor using all o f t h t experiment parnmtttrs mentioned above. except for the agi#ation
ratt, #howl that the results a r t as follows. The highest rpcciClc growth war at 0,082 hour-' at an aeration and
a~itacloar a t t of 0.5 vvm and 150 rpm. respectively. Based on the data of tht cultivation Irlneticn, the optlmum
coaditiona for tbt ftrmcntation in Biostat ZL-bioreictor is 1 vvm and 200 tpm of atntion and agitation, respectlvrly.
Tbc tMcltncy of substrate (Ypls) and of cell biomass (YI,,) to produce xylanase is 50.744 U.g-I mod 43.906 U.g-I,
rerptctivtly. T h t eMciency of substrate to cell production (Y*,,) is 1.178g.g-I. The liquid culttvatlon-mrdlrm h i s
non-Newtonian proptrtlts. Based on a mathematical model it i s Connd that the conulsttary ladex (k congtant)
mmd index o f liquid bthavlor (11 value) are 0.179 g.crn-'.8ccond-' and 0.3212. resptttivrly. Btcoust the value or
OcmO. the culture liquid i s categorized as pseudo plastic one. The Rbtynold number (NRc)
I s 6.9 x lff which indicates i t has turbulent cltrmcttristics. From a cmlcut~tioni t is louod that the power required
t o run a snitably slztd impeller i a 0.228 HP (Hornt Power) and the power ronsumption per unit volume i8
0.2265 HP.m4. A l l tbtsc valuer were u r t d Tor ncaling up xylanasc production ia the biortattor.
Key words: Bacillus pumilus, xylanase, production
Xylanase is a group o f extracellular enzymes which are
able to hydrolyze hemicelullosic materials into xylose and
xylo-oligosaccharides. Xylanases show great potential for
industrial applications mainly for the bioconversion of
lignocelluloses to sugar, ethanol, and other useful
substances,clarification of juices and wines, improving the
nutritional quality of silage and green feed and the de-inking
processes of waste papers (Viikari et a/. 200 1 ). The enzymes
have been grouped basedon the type of substrate hydro1ized.
Xylanases are categorized into three groups: P-xy losidase,
exoxylanase and endoxylanase. Endoxylanase i s the main
enzyme responsible for the cleavage of the linkages with in
the xylan backbone (Belfaquih ef al. 2002). Only a few
microorganisms are capabIe of producing xylanase
extraceluHarly. Some findings indicated that bacteria (Gilbert
and Hazlewood 1993; Sunna and Antranikian 1997), fungi
(lonukari et al. 2002; Kheng and Omar 200S), actinomycetes
(Ball and McCarthy 1989; Begg er at. 2001), and yeast
(Hnmva el ai, 1984;Liu a al. 1999) were capableof producing
xylanase, Xylanase-producing alkaliphillic bacteria can be
used as bleaching agent for paper processing (Ruiz-Arribas
et al. 1995).
%sent
address and *Corresponding author, Indonesian Center
for Agricultural Postharvest Research and Development, Pusat
Penelitian dan Pcngcrnbangan Pertanian. Jalan Tentara Pelajar
No 12, Bogor 16 1 14. Indonesia; Phone: +62-25 1-7 1 77064,
Fax: +62-251-321762, E-mail: rlch4n4@yahoo.co.id
Cultivation of xylanase-producing microorganisms in a
liquid medium has been applied widely in producing the
enyme. There are many advantages in using a liquid medium,
i.e. the type of component and composition of the medium
are easy to adjust to obtain the optimum conditions for
microbial growth; more efficient consumption of substrate
easier in adjusting the microorganism growth rate; and the
risk of contamination is less compared to using a solid
medium. However, the cultivation in liquid medium required
skills and expertise in operating a bioreactor for obtaining a
high level of enzyme production. By shaking the liquid
medium, it is expected that temperature, pH, oxygen,nutrient
supply as well as other environmental factors are
homogenous throughout the medium in ihe bioreactor.
The engineering on the processing of xylanase
production is based on the information obtained through
studying on the optimization process, the strategy on the
studying optimum level of substrate, and then the modeling
competency of the fermentation industry. All these cover
the studies on the cultivation kinetics based on three rates
i.e. the rates of biomass production, of substrateconsumption
and of enzyme production.
Production of xylanase from isolate bacteria capable of
utilizing xy Ian for -growth substrate as carbon source. In
general, the agro-residues such as wheat bran, sugar cane
bagasse*
cobs
paddy straw, rice husks* and cassava
wastes were found to be more suitable like xylan (Saurabh et
al. 200 1 ;Richana el a/.2004). Saha (2002) reported structure
The Process of Xylanase Production from
Baci~mpumilusRXAIII-5
NUR RICHANA1$*,TUN TEDJA IRAWADI', M. ANWAR NUR',
ILLAH SAILAH2, m KHASWAR SYAMSU2
IDepartment of Chemistry, F a d @ of Ydhemafics and Naiuml S d e n m
'DqmfmentofAgrolnhstdal TechnoIogy, F~acwflyof Agrolmrlrrsfrial Technology
Insiitri Pemnian Bogor, Darmaga Cartrprrs, Bogor 16680,Indonesiu
The optlmum conditions for the growth or Bsclllns pnmllus R X A I I I - 5 (a potential xylanist producer) were
nought. tbtst included trmpcraturc, pH, aeration. and agitation of the calturt batcb. Afterwards a mathematical
modtl bawd on the parameter of cultivation klnttics was formulattd. At the uamc time, the rheology o f the fluid
a d COT battcrlal crltlvatlon in a biortactor was studied. The data obtained was used for cstlmatlng the 'scaling
tip' of tnzyme production. The results of tbc study indicate thmt the optimum rondltion for proceraing I n 50 ml
E r l t n m t y t r flask a r t used temperature of 35 'C (308'K), pH 7, and an agitatlon rate or 140 rpm. The bigbest
xylama8c activity aod Its sprciflr activity arc 297.132 U,ml-I mod 655.32 U.g-'profitin, rtnptctlvtly. Subsequent
cxperimtntn In a blorcactor using all o f t h t experiment parnmtttrs mentioned above. except for the agi#ation
ratt, #howl that the results a r t as follows. The highest rpcciClc growth war at 0,082 hour-' at an aeration and
a~itacloar a t t of 0.5 vvm and 150 rpm. respectively. Based on the data of tht cultivation Irlneticn, the optlmum
coaditiona for tbt ftrmcntation in Biostat ZL-bioreictor is 1 vvm and 200 tpm of atntion and agitation, respectlvrly.
Tbc tMcltncy of substrate (Ypls) and of cell biomass (YI,,) to produce xylanase is 50.744 U.g-I mod 43.906 U.g-I,
rerptctivtly. T h t eMciency of substrate to cell production (Y*,,) is 1.178g.g-I. The liquid culttvatlon-mrdlrm h i s
non-Newtonian proptrtlts. Based on a mathematical model it i s Connd that the conulsttary ladex (k congtant)
mmd index o f liquid bthavlor (11 value) are 0.179 g.crn-'.8ccond-' and 0.3212. resptttivrly. Btcoust the value or
OcmO. the culture liquid i s categorized as pseudo plastic one. The Rbtynold number (NRc)
I s 6.9 x lff which indicates i t has turbulent cltrmcttristics. From a cmlcut~tioni t is louod that the power required
t o run a snitably slztd impeller i a 0.228 HP (Hornt Power) and the power ronsumption per unit volume i8
0.2265 HP.m4. A l l tbtsc valuer were u r t d Tor ncaling up xylanasc production ia the biortattor.
Key words: Bacillus pumilus, xylanase, production
Xylanase is a group o f extracellular enzymes which are
able to hydrolyze hemicelullosic materials into xylose and
xylo-oligosaccharides. Xylanases show great potential for
industrial applications mainly for the bioconversion of
lignocelluloses to sugar, ethanol, and other useful
substances,clarification of juices and wines, improving the
nutritional quality of silage and green feed and the de-inking
processes of waste papers (Viikari et a/. 200 1 ). The enzymes
have been grouped basedon the type of substrate hydro1ized.
Xylanases are categorized into three groups: P-xy losidase,
exoxylanase and endoxylanase. Endoxylanase i s the main
enzyme responsible for the cleavage of the linkages with in
the xylan backbone (Belfaquih ef al. 2002). Only a few
microorganisms are capabIe of producing xylanase
extraceluHarly. Some findings indicated that bacteria (Gilbert
and Hazlewood 1993; Sunna and Antranikian 1997), fungi
(lonukari et al. 2002; Kheng and Omar 200S), actinomycetes
(Ball and McCarthy 1989; Begg er at. 2001), and yeast
(Hnmva el ai, 1984;Liu a al. 1999) were capableof producing
xylanase, Xylanase-producing alkaliphillic bacteria can be
used as bleaching agent for paper processing (Ruiz-Arribas
et al. 1995).
%sent
address and *Corresponding author, Indonesian Center
for Agricultural Postharvest Research and Development, Pusat
Penelitian dan Pcngcrnbangan Pertanian. Jalan Tentara Pelajar
No 12, Bogor 16 1 14. Indonesia; Phone: +62-25 1-7 1 77064,
Fax: +62-251-321762, E-mail: rlch4n4@yahoo.co.id
Cultivation of xylanase-producing microorganisms in a
liquid medium has been applied widely in producing the
enyme. There are many advantages in using a liquid medium,
i.e. the type of component and composition of the medium
are easy to adjust to obtain the optimum conditions for
microbial growth; more efficient consumption of substrate
easier in adjusting the microorganism growth rate; and the
risk of contamination is less compared to using a solid
medium. However, the cultivation in liquid medium required
skills and expertise in operating a bioreactor for obtaining a
high level of enzyme production. By shaking the liquid
medium, it is expected that temperature, pH, oxygen,nutrient
supply as well as other environmental factors are
homogenous throughout the medium in ihe bioreactor.
The engineering on the processing of xylanase
production is based on the information obtained through
studying on the optimization process, the strategy on the
studying optimum level of substrate, and then the modeling
competency of the fermentation industry. All these cover
the studies on the cultivation kinetics based on three rates
i.e. the rates of biomass production, of substrateconsumption
and of enzyme production.
Production of xylanase from isolate bacteria capable of
utilizing xy Ian for -growth substrate as carbon source. In
general, the agro-residues such as wheat bran, sugar cane
bagasse*
cobs
paddy straw, rice husks* and cassava
wastes were found to be more suitable like xylan (Saurabh et
al. 200 1 ;Richana el a/.2004). Saha (2002) reported structure