OPTIMIZATION OF VARIOUS PARAMETERS FOR THE PRODUCTION OF

CELLULASE BY ASPERGILLUS NIGER

By:
Richa Singh
Shachi
Smriti Khandelwal
Tanvi Jain

DEPARTMENT OF BIOTECHNOLOGY
FACULTY OF ENGINEERING & TECHNOLOGY
R.B.S COLLEGE, BICHPURI CAMPUS, AGRA.
May 2009
OPTIMIZATION OF VARIOUS PARAMETERS FOR THE PRODUCTION OF
CELLULASE BY ASPERGILLUS NIGER
By:
Richa Singh
Shachi
Smriti Khandelwal
Tanvi Jain
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in partial fulfillment of the requirement for the degree of
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May 2009
CONTENTS
T"#-$/ Pa.$ *&.
DECLARATION55555555555555555555555555555....."
CERTIFICATE555555555555555555555555555555...6
AC7NO8LEDGEMENT55.55555555555555555555555....1
ABSTARCT55555555555555555555555555555555.-
LIST OF ABBREVIATIONS5555555555555555555555555!
LIST OF TABLES55555555555555555555555555555..*
LIST OF FIGURES55555555555555555555555555555.&
CHAPTER 9: INTRODUCTION5555555555555555555555......9
CHAPTER 2: REVIE8 OF LITRATURE5555555555555555555..;
2.1 eneral !"pect"#############################....$
2.2 Type" of %ellula"e############################$
2.& 'echani"m of %elluloly"i"########################...(
2.) %ellula"e *roducing 'icroorgani"m####################....+
2.).1 ,ungi##############################...+
2.).2 Bacteria.-#############################...
2./ Su0"trate 1"ed for the production of %ellula"e################.12
2.$ 'ethod of *roduction##########################11
2.$.1 Solid State ,ermentation#####################.#..12
2.$.23ifferent Su0"trate" for Solid State ,ermentation#############.12
2.$.& Su0merged ,ermentation######################...1&
2.$.) 3ifferent Su0"trate for Su0merged ,ermentation#############.1)
2.(4n5yme !""ay 'ethod##########################1/
2.(.13initro"alicyclic!cid 'ethod#####################.1/
2.(.2 ,ilter *aper !""ay 'ethod######################1$
2.+ 6ndu"trial !pplication of %ellula"e #####################1(
2.+.1 !pplication of %ellula"e to %ontrol 6ndu"trial Slime##########...#.1(
2.+.2 !pplication of %ellula"e in the Sterility Te"t for ,ortified *rocaine *enecilline .1(
2.+.& %ellula"e 1"edfor Starch *roduction##################.1+
2.+.)%ellula"e 1"ed for ,uel *roduction###################1+
CHAPTER<:MATERIALS AND METHODS555555555555555555.99
&.1'aterial"###############################..22
&.1.1 %hemical 1"ed###########################22
&.1.2 'icroorgani"m"##########################..21
&.1.& 4quipment"############################.21
&.1.) 'edia 1"ed############################21
&.2 'ethod"###############################..2&
&.2.1 'aintenance of Stoc7 %ulture#####################2&
&.2.2 Su0"trate 1"ed for *roduction of %ellula"e################2&
&.2.& *retreatment of Su0"trate#####.........................................................................2)
&.2.) %ellula"e *roduction 0y Su0merged ,ermentation#############..2$
&.2./ %ellula"e *roduction 0y Solid State ,ermentation#############..2(
&.2.$ 4n5yme !""ay###########################..2(
&.2.$.1 *reparation of luco"e Standard %urve##############..2+
CHAPTER =: RESULT AND DISCUSSION555555555555555555.<9
).1 Selection of Su0"trate for *roduction of %ellula"e###############.&2
).2 Selection of ,ermentation %ondition" for *roduction of %ellula"e#########&/
).& 8ptimi5ation of Temperature for *roduction of %ellula"e############.)2
).) 8ptimi5ation of ,ermentation %ondition" for *roduction of %ellula"e#######)&
CHAPTER ;: CONCLUSION55555555555555555555..5555.=>
CHAPTER >:BIBLIOGRAPHY 5555555555555555555.5555..=?
DECLARATION
9e here0y declare that thi" "u0mi""ion i" our own wor7 and that: to the 0e"t of our
7nowledge and 0elief: it contain" no material previou"ly pu0li"hed or written 0y another per"on
nor material which to a "u0"tantial e;tent ha" 0een accepted for the award of any other degree or
diploma of the univer"ity or other in"titute of higher learning: e;cept where due ac7nowledgment
ha" 0een made in the te;t.
Na!$ R&-- N&. S".*a#u)$
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S'a,'" 0;00=;=022
S!)"#" 7'a*%$-2a- 0;00=;=02=
Ta*0" @a"* 0;00=;=02?
Da#$:
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CERTIFICATE
Thi" i" to certify that *ro<ect Report entitled =8ptimi5ation of variou" parameter" for the
production of cellula"e 0y Aspergillus niger>: which i" "u0mitted 0y Richa Singh:Shachi:Smriti
Khandelwal:Tanvi Jain in partial fulfillment of the requirement for the award of degree B. Tech.
in Biotechnology from 1. *. Technical 1niver"ity: i" a record of the candidate own wor7 carried
out 0y him under my "upervi"ion. The matter em0odied in thi" the"i" i" original and ha" not 0een
"u0mitted for the award of any other degree.


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B",'(u)" Ca!(u/, A.)a

364
AC7NO8LEDGEMENT
6t give" u" a great "en"e of plea"ure to pre"ent the report of the B. Tech *ro<ect underta7en during
,inal ?ear. 9e owe "pecial de0t of gratitude to lecturer @.K. Sriva"tava: 3epartment of BioA
technology: for hi" con"tant "upport and guidance throughout the cour"e of our wor7. Bi"
"incerity: thoroughne"" and per"everance have 0een a con"tant "ource of in"piration for u". 6t i"
only hi" cogni5ant effort" that our endeavor" have "een light of the day.
9e al"o ta7e the opportunity to ac7nowledge the contri0ution of 4r. !"hi"h
Shu7la:Cecturer of Biotechnology department for hi" full "upport and a""i"tance during the
development of the pro<ect.
9e would al"o li7e to appreciate the contri0ution of 4r. 'ano< Kumar: and 4r. Sha7ti
Katiyar Cecturer of Biotechnology department for their 7ind a""i"tance and cooperation during
the development of our pro<ect. 9e are al"o grateful to the technical "taff without who "e
cooperation we would have not 0een a0le to complete our pro<ect. 9e would al"o than7 our
friend" for their contri0ution in the completion of the pro<ect.
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Da#$:

314
ABSTRACT
%ellula"e" are a group of hydrolytic en5yme capa0le of degrading cellulo"e to the "maller
gluco"e unit". The goal of the pre"ent "tudy wa" to inve"tigate the cellula"e en5yme production
a0ility of A.niger again"t cellulo"ic material" "uch a" 9heat 0ran: "ugarcane: Baga""e: *aper:
cotton and "elect the 0e"t %ellula"e en5yme producing "u0"trate . ma;imum en5yme activity
o0tained wa" Baga""e 1..(2Dmol-min-ml.
6t wa" u"ed for optimi5ation of temperature which wa" o0tained at &/E% with a ma;imum
activity of 1../+Dmol-min-ml.
Time wa" al"o optimi5ed with a ma;imum en5yme activity of 1..(2 Dmol-min-ml at 12 day"
interval time.

3-4

3S!4
L"/# &+ Ta-$/
TABLE 9: 3ifferent ,ungi wor7ed upon 0y different author" for %ellula"e production
TABLE 2: 3ifferent Bacteria wor7ed upon 0y different author" for %ellula"e production
TABLE <: 3ifferent Su0"trate wor7ed upon 0y different author" for %ellula"e production
TABLE =: !0"or0ance of luco"e Standard %urve
TABLE ;: 4n5yme !ctivity found for different Su0"trate"
TABLE >3a4: 4n5yme !ctivity found for different fermentation condition at &2E%
TABLE >34: 4n5yme !ctivity found for different fermentation condition at &/E%
TABLE A: 4n5yme !ctivity found for different Temperature"
TABLE ?: 4n5yme !ctivity found for different day" interval


3*4
L"/# &+ F".u)$/
F".u)$ 9: *repared "lant of Su0 cultured A.niger
F".u)$ 23a4: *retreated "u0"trate" Baga""e FleftG: 9heat Bran FrightG
F".u)$ 234: *retreated "u0"trate" %otton FleftG : *aper FrightG
F".u)$ <: luco"e Standard %urve
F".u)$ =: 3iagram "howing !0"or0ance for 3ifferent Su0"trate"
F".u)$ ;3a4: 3iagram "howing !0"or0ance for 3ifferent ,ermentation condition at &2E%
F".u)$ ;34: 3iagram "howing !0"or0ance for 3ifferent ,ermentation condition at &/E%
F".u)$ >: 3iagram "howing !0"or0ance at 3ifferent Temperature
F".u)$ A: 3iagram "howing !0"or0ance at 3ifferent ,ermentation time Fday"G




3&4
INTRODUCTION
REVIE8 OF LITERATURE
MATERIALS AND
METHODS
CONCLUSION
BIBLIOGRAPHY