Post on 04-Jun-2018
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 1/7
E L S E V I E R
Process iochemistry Vol . 33, No. 3 , pp. 323-3 29, 1998© 1998 P ub l i s he d by E l s e v ie r S c i e nc e L t d
A l l r i gh t s r e se r ve d . P r i n t e d i n G r e a t B r i t a in0032-9592/98 19.00 + 0.00
P I I : S 0 0 3 2 - 9 5 9 2 9 7 ) 0 0 0 8 6 - 1
ranberry process ing waste for sol id s tatefungal inoculant product ion
Zuo xing Zh eng Ka lidas Shetty*
De par tm ent o f Food Science, Universi ty of M assachuset ts , Am herst , MA 01003, USA
(Received 7 July 1997; revised version received 28 Augu st 1997; accepted 1 Septem ber 1997)
bstract
Cran berry pom ace is a primary by-product of the trad it ional cranberry juice processing industry and i tsdisposal presents economic and environmental problems. Microbial conversion of cranberry pomace intovarious value-added products is a practical approach for solving such disposal problems. The presentresearch was undertaken to test the growth of several agriculturally and industrial ly important fungi oncranberry pomace substrate through solid-state fermentation. Fungi, such as Trichoderma viride If-26, Trichoderma harzianum ATC C 24274 , and Trichoderma pseudokoningii ATCC 26801, a novel polymeric dyedecolorizing PeniciUium isolate, and a food-grade Rhizopus st rain i solated f rom Tempeh, that produceindustrially imp ortan t extracellular enzym es were grown on a cranb erry pomace-based me dium a t 25°C for4 days. The glucosamine content o f the h eterogeneous ferme nted m ixture was a good indicator of fungalgrowth. Th e maximum growth o f al l fungi was establi shed on cranberry poma ce su pplemen ted wi th 0 .05 go f C a C O 3 , 2 0 ml o f water, and 0 05 g of N H 4 N O 3 o r 0 2 ml o f fish protein hydrolysate pe r gram of pomace.It was concluded that bioconversion of cranberry processing waste by industrial ly beneficial fungi throughsolid-state ferm enta tion was feasible. This po tential can be coup led with the uti lization of fish processingwaste as an organ ic nitrogen source to develop m utually com plem entary produ cts benefi t ing bo th the fisheryand c ranbe rry processing industries. © 1998 Elsevier Science Ltd
Keywords: cranb erry waste, fungal inoculants, glucosamine co ntent, solid-state ferm entation .
I n t r oduc t i on
A p p r o x i m a t e l y 4 2 0 m i l l i o n p o u n d s o f c r a n b e r r i e s a r e
p r o d u c e d a n n u a l l y i n t h e U S A , a b o u t 9 0 % o f w h i c h
a r e u s e d f o r p r o c e s s i n g p u r p o s e s [ 1 ]. T h e p r i m a r y
b y - p r o d u c t o f t r a d i t i o n a l c r a n b e r r y j u i c e p r o c e s s i n g i s
c r a n b e r r y p o m a c e . I t c o n si s ts o f t h e p r o c e s s e d s k i n s ,
s e e d s a n d s t e m s , a n d c o n s t i t u t e s a b o u t 5 % o f t h e w e t
w e i g h t o f t h e o r i g i n a l f r u it . F r e s h l y p r e s s e d c r a n b e r r y
p o m a c e c o n t a i n s a l a r g e a m o u n t o f i n s o l u b l e c a r b o -
h y d r a t e s w i t h s m a l l a m o u n t s o f p r o t e i n , m i n e r a l s a n d
s o m e r e m a i n i n g j u ic e w i t h s u g a r s a n d o t h e r s o l u bl e
s u b s t a n c e s . O w i n g t o i t s h i g h m o i s t u r e c o n t e n t , f r e s h l y
p r e s s e d c r a n b e r r y p o m a c e i s s u s c e p t i b l e t o r a p i d
m i c r o b i a l g r o w t h . L i k e o t h e r f r u i t p r o c e s s i n g w a s t e s ,
s u c h a s a p p l e p o m a c e [ 2 ] , g r a p e p o m a c e [ 3 ] , t o m a t o
pomace [4 ] , c i t ru s was t e [5 ] , p i neapp l e was t e [6 ] ,
*To whom correspondence should be addressed. Tel . : (+1)413 545 10 22 ; fa x : (+1) 413 545 12 62 ; e-mail :kalidas@foodsci .umass.edu
323
o r a n g e w a s t e [ 7 ] , s u g a r - c a n e p r e s s m u d [ 8 ] , a n d k i w i -
f r u i t p e e l w a s t e [ 9] , c r a n b e r r y p r o c e s s i n g w a s t e i s c o m -
m o n l y u s e d a s a n i m a l f e e d o r f e r t i l i z e r . H o w e v e r , i t s
v a l u e a s a n i m a l f e e d i s v e r y l i m i t e d b e c a u s e o f i t s l o w
p r o t e i n c o n t e n t a n d i t s u s e a s f e r t i l i z e r m a y n o t b e
e c o n o m i c a l l y c o m p e t i ti v e . F u r t h e r , d i r e c t d is p o s a l o f
pomace was t e t o so i l o r i n a l and f i l l poses s i gn i f i can t
e n v i r o n m e n t a l p r o b l e m s . T h u s , t h e e x p l o r a t i o n o f
n o v e l u s e s f o r c r a n b e r r y w a s t e i s n e e d e d .
B i o l o g i c a l c o n v e r s i o n o f f r u i t p r o c e s s i n g w a s t e s i n t o
v a r i o u s v a l u e - a d d e d p r o d u c t s t h r o u g h s o l i d - s t a t e f e r -
m e n t a t i o n ( S S F ) h a s b e e n o f m a j o r i n t e re s t t o m a n y
l a b o r a t o r i e s a r o u n d t h e w o r l d . S S F d e a l s w i t h t h e u t i l i -
z a t i o n o f w a t e r - i n s o l u b l e m a t e r i a l s f o r m i c r o b i a l
g r o w t h a n d m e t a b o l i s m , a n d i t is u s u a ll y c a r r i e d o u t i n
s o l id o r s e m i - s o l i d s y s te m s i n t h e n e a r a b s e n c e o f f r e e
w a t e r o r r e d u c e d w a t e r c o n t e n t c o m p a r e d w i t h s u b -
m e r g e d f e r m e n t a t i o n [ 1 0 ] . M a n y o f t h e p o t e n t i a l
p r o d u c t s f r o m f r u i t p o m a c e w a s t e s h a v e b e e nd e v e l o p e d u s i n g t h e S S F t e c h n i q u e , a n d s u c h p r o d u c t s
i nc l ude e t hano l [11 ] , met hane [7 ] , l ac t i c ac i d [8 ] , c i t r i c
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 2/7
324 Bioconversion of cranberry processing waste
acid [6], protein [5], mushrooms [12], enzyme [13], and
fo o d i n g re d i en t [ 3 ] . H o w e v e r , n o r e p o r t h a s b e e n
foun d dea l ing wi th the u t i l iza t ion o f c ranberry p ro -
cess ing was te .
Trichoderma i s no tab ly capab le o f p roducing var ious
p o l y s a c c h a r i d e -d e g ra d i n g e n z y m e s w h i c h e n a b l e i t t o
g ro w o n c r a n b e r ry p o m a c e s u b s t r a te . I t h a s b e e n
re p o r t e d t h a t s o m e Trichoderma spec ies a re wide ly
used to p roduce var ious indus t r i a l ly impor tan t enzymes
[14-16] . Trichoderma spec ies can a l so be e f fec t ive as
b io log ica l con t ro l agen ts aga ins t pa thogen ic o rgan isms
which usua l ly cause many p lan t roo t d i seases [17 ] . Tr / -
c h o d e r ma h a r z i a n u m a l so has the ab i l i ty to degrade
organoch lo r ine pes t i c ides , such as DDT, d ie ld r in ,
e n d o s u l f a n , p e n t a c h l o ro n i t ro b e n z e n e , a n d p e n t a c h l o r -
opheno l , and hence has po ten t ia l app l ica t ions fo r b io -
remed ia t ion [18 ] .
Severa l fungal spec ies be long ing to the genus Rhi -
z o p u s have been used in SSF fo r severa l cen tu r ies ,e s p e c i a l l y i n A s i a fo r p r e p a r i n g m a n y f e rm e n t e d fo o d -
stuffs . R h i z o p u s no t on ly enhances the d iges t ib i l i ty and
pro te in c on ten t o f foods tu f fs , bu t a l so p reven ts the
fo rmat ion o f tox ic subs tances such as a f la tox in B1 .
S o m e R h i z o p u s spec ies can a l so p roduce an t i -carc ino -
genic substances and ant ibiot ics [19,20]. A s train of
Rhizopus oligosporus, w h i c h w a s i s o l a t e d f ro m c o m m e r -
c ia l Tempeh in ou r l abora to ry , was used in th i s s tudy in
o rd e r t o d e v e l o p a p o t e n t i a l p ro t e i n - e n h a n c e d v a l u e -
a d d e d p ro d u c t f r o m c r a n b e r ry p o m a c e fo r u s e a s
an imal feed .
An o the r fun gus used in th i s s tudy is a novel Pen-icillium i so la te wh ich i s capab le o f deco lo r iz ing the
po lymer ic dyes Po ly R-478 and Po ly S-119 in l iqu id
med ia [21 ] . Th is i so la te has po ten t ia l app l ica t ions in
b i o r e m e d i a t i o n o f a ro m a t i c p o l l u t a n t s s i n ce i t c o u ld b e
u s e d t o r e m o v e s o m e d y e s t u ff s f r o m d y e -c o n t a m i n a t e d
water or soi l .
F i sh o f fa l i s a majo r f i shery by -p roduct wh ich i s
u s u a l l y d i s p o s e d o f o n l a n d o r o f f s h o re a s w a s t e e v e ry
year. Since i t has a h igh ni t rogen content [22], i ts acid
h y d ro l y s a t e c o u l d b e s u p p l e m e n t e d t o c r a n b e r ry
p o m a c e m e d i u m t o e n r i c h t h e o rg a n i c n i t r o g e n fo r
fungal g rowth . The ob jec t ive o f th is research w as to
d e v e l o p n o v e l a p p ro a c h e s t o u t i l i z e c r a n b e r ry p o m a c e ,coup led wi th u t i l i za t ion o f f i shery was te , to genera te
va lue-added p roduct s , l ike microb ia l inocu lan t s , u s ing
t h e b e n e f i c i a l f u n g i m e n t i o n e d a b o v e . C ra n b e r ry
p o m a c e c o u l d n o t o n l y s e rv e a s a n e x c e l l e n t c a rb o n
source , bu t in add i t ion i t cou ld be used as an o rgan ic
car r ie r fo r fungal inocu lan t s fo r food , ag r icu l tu ra l and
env i ronmen ta l app l ica t ions .
a t e r ia l s a n d m e t h o d s
Microorgan isms
T. viride IF-26, T. h a r z i a n u m ATCC 24274 , and T. pseu-
dokoningii A TC C 2 6 8 0 1 w e re o b t a i n e d f ro m t h e
A m e r i c a n Ty p e C u l t u r e C o l l e c t i o n (R o c k v i l l e , M D ) ; a
Penicillium sp . ATCC 74414 tha t deco lo r ized po lymer ic
dyes was i so la ted in ou r l a bora to ry [21] ; a s tra in o f
Rhizopus oligosporus w a s i s o l a t e d f ro m u n p a s t e u r i z e d
Te m p e h p ro d u c t . Th e Te m p e h p ro d u c t w a s k i n d l y p ro -
v i d e d b y L i f e -L i f e F o o d s C o . , G re e n f i e ld , M A .
Media and cul t ivat ion condit ion
Th e m i c ro o rg a n i s m s w e re m a i n t a i n e d o n p o t a t o d e x -
t rose agar (PDA) s lan t s and pe t r i p la tes a t 4 °C and
subcu l tu red mon th ly . Al l fung i were cu l tu red a t room
t e m p e ra t u r e fo r 7 d a y s b e fo re u s e . 1 2 5 m l E r l e n m e y e r
f lasks con ta in ing 10 g o f c ranbe rry pom ace , 0 .5 g o f
CaCO 3, 20 ml water , and 0 5 g o f NH aNO 3 o r 2 ml f ish
p ro t e i n h y d ro l y s a t e (F P H ) a s t h e s u p p l e m e n t a l
n i t rogen source were used fo r SSF. The f resh ly p ressed
cranberry pomace was ob ta ined f rom Very f ine , Inc . ,
W e s t fo rd , M A , a n d w a s d r i e d a n d g ro u n d a n d s t o r e din a re f r igera to r b efo r e use . The wate r con ten t o f c ran -
berry pomace used in the exper imen t was 5 .8% (w/w,
w e t b a s i s ) . F P H w a s o b t a i n e d f ro m O c e a n C re s t
(Glouces te r , MA) as her r ing was te con ta in ing
0 .6575 g m1-1 o f so lub le so l ids . The spores f rom one
PDA p la te were inocu la ted in to abou t 20 f l asks . The
f lasks were incubated a t 25°C fo r 4 days . The cu l tiva-
t ion o f a l l fung i was a l so ex t rapo la ted fo r 100 g o f
c r a n b e r ry p o m a c e w i t h p ro p o r t i o n a l a d d i t i on o f o t h e r
supp lem en ts ca lcu la ted f ro m the 10 g l eve l.
Protein assay
100 ml o f d i s t il l ed wa ter was ad ded in to the f ung us -
pomace-con ta in ing f l asks and the cu l tu re was homoge-
n ized us ing a War ing b lender , then cen t r i fuged a t
1500g fo r 15 min . The supern a tan t was used fo r p ro te in
assay . So lub le p ro te in was de te rmined us ing a commer-
c ia l as say k i t (B io -R ad Pro te in Assay Ki t I I , B io -Ra d
La b o ra t o ry , H e rc u l e s , C A ) w i t h b o v i n e s e ru m a l b u m i n
a s s t a n d a rd , a c c o rd i n g t o t h e p ro c e d u re d e s c r i b e d b y
Bra dfo rd [23] . The so lub le p ro te in p rod uce d by fungal
s t ra i n s i n t h e c r a n b e r ry p o m a c e m e d i u m w a s e x p re s se d
as mi l l ig rams per g ra m o f pom ace (o r ig ina l d ry
weigh t ) .
Mo isture content M C) and wa ter act ivi ty determination
T h e M C o f cr a n b e rr y p o m a c e m e d i u m w a s d e t e r m i n e d
b y m e a s u r in g b o t h t h e w e t w e i g h t a n d d ry w e i g h t o f
the sample . Af te r measur ing the wet weigh t , the
samp le was d r ied in an ov en a t 105°C fo r 2 days , o r
un t i l the weigh t was cons tan t , befo re reco rd ing the d ry
w e i g h t. Th e w a t e r a c ti v it y a w o f th e c r a n b e r ry p o m a c e
m e d i u m w a s d e t e rm i n e d a c c o rd i n g t o t h e m e t h o d
d e s c r i b e d b y M c C u n e et al. [24 ] . A re ference mater ia l
(c i rc le o f f i l t e r paper) o f known so rp t ion i so therm wasob ta in ed by equ i l ib ra t ing fo r 24 h to each o f s ix sa lt
s lu shes and was equ i l ib ra ted fo r 24 h to the sample ; the
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 3/7
Z. Zheng and K. Shetty 325
water activity of the equilibrated filter paper was cal-
culated using the linear equation of the isotherm curve
from its MC, which was determined by measuring the
weight gain of the filter paper during equilibration. The
water activity of the sample was equal to that of the
equilibrated filter paper. The relationship between MC
(wet basis) and aw of the sample was expressed by the
equation
MC = A + B log (1 - a w
where, at 25°C,A = 0.0033 and B = -0.1155.
Glucosamine assay
The glucosamine content of the fermented culture
mixture consisting of fungal mycelia and cranberry
pomace medium was used to estimate fungal biomass
during the SSF as the growth indicator of Trichoderma,
Rhizopus , and Penicillium strains. It was determined bythe modified method of Sakurai et al. [25]. After culti-
vation, the culture mixture in each flask was mixed with
100ml of distilled water. The mixture was then
homogenized using a Waring blender. A 1 ml suspen-
sion of homogen ized sample was mixed with 2 ml of
H2504 (98 ) in a test tube. After standing for 24 h at
25°C, it was diluted with 47 ml of water and autoclaved
at 120°C for 1 h. The hydrolysate was then neutra lized
with NaOH to pH7-0 and diluted to 100ml, from
which 0.5 ml was mixed with 0.5 ml of NaNO2 (5%)
and 0-5 ml of KHSO4 (5%) in a centrifuge tube. After
shaking occasionally for 15 min, it was centrifuged at1500g for 2 min. 0.6 ml of supernatant was then mixed
with 0 2 ml of NHaSO3NH2 (12 5%) and shaken for
3 min. To the mixture, 0.2 ml of 3-methyl-2-benzothia-
zolinone hydrazone hydrochloride (MBTH, 0.5%, pre-
pared daily) was added and then the mixture was
boiled for 3 min. The reaction mixture was immediately
cooled to room temperature following boiling and
0.2 ml of FeCI3 (0 5%, prepared within 3 days) was
added. After standing for 30min, the absorbance at
650nm was measured spectrophotometrically. The
glucosamine content was calculated as milligrams per
gram of pomace (original dry weight) according to the
standard curve.
e s u l t s a n d d i s c u s s i o n
The effect of Ca C0 3 supplementation on the growth of
selected fungi
Since the cranberry pomace contains organic acids, the
pH of cranberry-pomace-based medium was relatively
low (approximately 3.0 to 3-2) and hence inhibited the
growth of fungi. In order to obtain the maximal
growth, it was necessary, therefore, to neutralize the
medium before inoculation. In general, most fungi areable to grow in a pH range of neutral to slightly acidic.
Considering the desired pH range, CaCO3 was con-
8 .
0
i i i i i i i
1 2 3 4 5 6 7 8 9 1
CaC03 g/ lOOg pomace)
Fig. l. The effect of CaCO3addition on the growth of Tricho-derma, Rhizopus and Penicillium strains on cranberry pomace.
sidered as an ideal neutralizer because it can increase
the pH of cranberry pomace medium to 5.8-7.0 and it
is inexpensive for large-scale use. Again, Trichoderma
viride , Rhizopus isolate, and Penicillium G-1 strains
were used to examine the effect of CaCO3 addition in
cranberry pomace on fungal growth using 2 mlg
water and 0 .05 g NH4NO3. It was demonstra ted that
cranberry pomace medium supplemented with
0.04-0.05 g of CaCO3 per gram of pomace would be
satisfactory for the growth of all three selected fungi
(Fig. 1).
The effect o f water addit ion in cranberry pom ace on the
growth of se lected fungi
The moisture level of the medium is a critical factor
influencing the growth of fungi in SSF. In general, a
higher moisture level results in decreased porosity or
intracellular spaces, lower oxygen diffusion and gas
exchange and enhanced formation of aerial mycelium.
In contrast, a low moisture level will lead to decreased
substrate swelling and decreased microbial growth. In
this study, partially dried and ground cranberry pomace
with an MC of 5 8% (w/w) was used. Since it had very
low water content, additional water was required toincrease the moisture level of the medium. To deter-
mine the optimal amount of water for maximum
growth, the growth of T. viride, T. pseud okonin gii, T.
harzianum, Penicil l ium G-l, and the Tempeh Rhizopus
isolate were examined using the basal cranberry
pomace medium with inorganic nitrogen as described
in the Material and methods section with varying water
contents. The soluble protein produced by various
fungi in the pomace medium was measured as the indi-
cator of growth (Fig. 2). All fungi tested in the experi-
ment grew very poorly if no additional water was
added to the medium; they exhibited very similargrowth patterns as the amount of water supplemented
to the medium increased up to 4mlg -~ before
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 4/7
326 Bioconve rsion of cranberry processing waste
0
0
¢ / / . / o - *- r . ~,, ,~o,,m
l
100 200 300 400 500
E
g
a 1
H20 ml/100g pomace)
Fig. 2. The effect of water addition in cranberry pomacemedium on the growth of Trichoderma Penicillium and Rhi-zopus strains.
declining slightly at higher water addition
(4.5-5-0 ml g-I). Addition of about 2 ml of water per
gram of cranberry pomace was sufficient for the
maximum growth of all selected fungal strains.
The relat ionship between M C a nd w ater activity o f
cranberry pom ace med ium
A better way of expressing the MC of cranberry
pomace medium is the water activity aw, which indi-
cates the availability of water for the growth of fungi.
The MC and water activity of the cranberry pomace
medium corresponding to each level of water supple-
mented to the medium were determined using the
methods described in the Materials and methods
section. It appeared that the optimal MC in the
pomace for the growth of all selected fungi was about
67 (wet basis), whereas the corresponding optimal
water activity was 0.99 (Table 1).
Bioma ss es t ima t ion
In SSFs, one of the most important problems encoun-
tered is the biomass measurement. Unlike submergedcultivation, fungal mycelia are intimately bound to the
Table 1. The effect of water addition on the MC and aw ofcranberry pomace medium
Water addition
to the medium(ml/g pomace)
MC in the pomace(%, wet basis)
w
0 5 79 0-46431 51 49 0-97472 67 34 0 99043 75-38 0 9938
4 80-05 0-99455 83 50 0-9960
solid matrix and cannot be quantitatively separated
from it; so, direct measurement of fungal biomass is
impossible. Many authors have described methods of
indirect biomass estimation including measuring: (a)
fungal cell constituents, such as ergosterol, nucleic
acids, protein, nitrogen and chitin; (b) primary metabo-
lites, such as CO2, ATP, or enzymatic activity; (3)
nutrient consumption [26, 27].
As glucosamine is an essential and stable compo-
nent in chitin of mycelial cell walls, the glucosamine
content seems to be a useful parameter for the estima-
tion of the total sum of the growing mycelium and its
changes may correspond to the development of the
mycelium, although the values cannot be converted to
mycelial weight quantitatively [25]. Roche et al. [27]
reported that a linear correlation existed between
cumulative biomass and cumulative glucosamine of fila-
mentous fungi during SSF. Desgranges et al. [26] also
suggested that the glucosamine measurement gave agood indication of fungal biomass development, but
the biomass indicator could only be used to compare
different media having the same constituents, even if
the C/N ratios were different.
In our study, two methods for biomass estimation
were compared when additional nitrogen source was
added to the medium. In the case when NH4NO3 was
supplemented as a nitrogen source, as shown in Figs 3
and 4, the measurement of soluble protein content was
consistent with the measurement of glucosamine
content, as both gave a similar growth pattern. Under
such conditions, therefore, either soluble protein or
glucosamine content could be used as the growth indi-
cator of the selected strains in SSF.
The e f fec t o f NH 4N 03 on the growth o f selec ted fung i
An important indicator of nutritional regulation of
growth in SSF is the C/N ratio. The optimal C/N ratio
Eg
J
n
i . - . , - -~
1
0 2 4 6 8 10
NI-14NOa g/100g Ix)mace)
Fig. 3. The effect of NH4NO3 addition on the growth ofTrichoderma Rhizopus and Penicillium strains on cranberrypomace.
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 5/7
Z. Zheng and K. Shetty 327
10
E8
E
(.9/ --X--R/~o/x~
- - - l . -Pe t~
2 4 6 8 10
NH4NOa (g/100g pomace)
Fig. 4. The effect of NH4NO suppleme ntat ion on the glucos-amine product ion by Trichoderma Rhizopus and Penicilliumstrains grown o n cranb erry pomace.
v a r i e s i n a w i d e r a n g e f r o m 1 0 t o 1 0 0 o r h i g h e r i n
v a r i o u s S S F p r o c e s s e s , b u t t h e a v a i l a bi l it y o f C a n d N
c a n b e m o r e i m p o r t a n t t h a n t h e r a t i o . I n m o s t S S F
s y s t e m s , t h e c a r b o n s o u r c e c o m e s f r o m t h e n a t u r a l
s o l u b l e a n d i n s o l u b l e c a r b o h y d r a t e s w h i l e t h e n i t r o g e n
s o u r c e i s a d d e d . C r a n b e r r y p o m a c e c o n t a i n s h i g h e r
c a r b o h y d r a t e s ( ~ 5 0 o n a d r y w e i g h t b a s is ) , s o m e o f
w h i c h a r e f e r m e n t a b l e a n d c o u l d b e u s e d a s a c a r b o n
s o u r c e , b u t a r e l a t i v e l y l o w c o n t e n t o f n i t r o g e n s o u r c e
a v a i l a b le f o r f u n g i b e c a m e t h e l i m i t in g f a c t o r f o r f u n g a lg r o w t h . T h e r e f o r e , i n o r d e r t o o b t a i n t h e o p t i m a l
g r o w t h o f fu n g i o n c r a n b e r r y p o m a c e , s u p p l e m e n t a t i o n
o f n i t r o g e n s o u r c e w a s n e e d e d . T w o k i n d s o f n i t r o g e n
s o u r c e , N H a N O 3 a n d F P H , w e r e u s e d i n t h i s s t u d y .
T h e g r o w t h o f T . v ir ide Rhizopus i so l a t e , and Peni-
cillium G - 1 w a s d e t e r m i n e d b y m e a s u r i n g b o t h t h e
s o l u b l e p r o t e i n c o n t e n t a n d g l u c o s a m i n e c o n t e n t o f
f e r m e n t e d c u l t u r e . T h e e f f e c t o f N H 4 N O 3 o n t h e
g r o w t h o f t h e t h r e e s e l e c t e d f u n g i i s s h o w n i n F ig . 3 ( i n
t e r m s o f s o l u b l e p r o t e i n p r o d u c t i o n ) a n d F i g . 4 ( i n
t e r m s o f g l u c o s a m i n e c o n t e n t ) . A d d i t i o n o f a b o u t
0 - 0 5 g o f N H 4 N O 3 p e r g r a m o f p o m a c e g a v e th e
m a x i m a l g r o w t h o f a l l s e l e c t e d f u n g a l s p e c i e s ( F i g s 3
and 4 ) .
The effect o f FP H on the growth o f selected fung i
F i s h o f f a l i s a n o t h e r i m p o r t a n t f o o d p r o c e s s i n g w a s t e ,
a n d i t s d i s p o s a l h a s n o t y e t b e e n s o l v e d s a t i s f a c t o r i l y
[ 2 2 ] . B y a c o m b i n a t i o n o f p a p a i n a n d a c i d h y d r o l y s i s ,
t h e f i s h er y b y -p r o d u c t w a s c o n v e r t e d i n t o F P H i n
w h i c h a h i g h c o n c e n t r a t i o n o f n u t r i e n t s , s u c h a s
n i t ro g e n , w a s e x p e c t e d t o e n h a n c e t h e f u n g a l g ro w t h
o n c r a n b e r r y p o m a c e m e d i u m . M a r t i n a n d C h i n t a l a p a t i[ 2 2 ] d e m o n s t r a t e d t h a t t h e g r o w t h o f Scytalidium acid-
ophi lum o n a c i d p e a t h y d r o l y s a t e w a s e n h a n c e d w h e n i t
g r e w i n f is h o f f a l - p e a t c o m p o s t . W e p r o p o s e d t h a t
F P H c o u l d b e a n a l t e r n a t i v e n i t r o g e n s o u r c e f o r c r a n -
b e r r y p o m a c e m e d i u m e n r ic h m e n t .
W e t e s t e d t h e e f f e c t o f s u p p l e m e n t a t i o n o f h e r r i n g
F P H o n t h e g r o w t h o f Trichoderma viride Rhizopus
i so l a t e , and Penicillium G - 1 s t r a i n s o n c r a n b e r r y
p o m a c e m e d i u m . F u n g a l g r o w t h w a s e x p e c t e d t o b e
e x p r e s s e d i n t e r m s o f b o t h s o l u b l e p r o t e i n a n d g l u c o s -
a m i n e c o n t e n t s . H o w e v e r , i t w a s p r o b l e m a t i c t o u s e
s o l u b l e p r o t e i n c o n t e n t a s t h e g r o w t h i n d i c a t o r b e c a u s e
t h e p a r t i a l l y h y d r o l y z e d p r o d u c t s o f f is h p r o t e i n , s u c h
a s p o l y p e p t i d e s i n h e r r i n g F P H s u p p l e m e n t e d i n t h e
m e d i u m , c o n t r i b u t e d t o t h e t o t a l s o l u b l e p r o t e i n
c o n t e n t o f t h e f e r m e n t e d m e d i u m . T h i s w o u l d e x p l a i n
t h e m i s l e a d i n g a n d c o n f u s i n g r e s u l t t h a t t h e s o l u b l e
p r o t e i n w a s c o n s t a n t l y i n c r e a s i n g a s t h e l e v e l o f F P H
s u p p l e m e n t e d t o t h e m e d i u m i n c r e a s e d ( d a t a n o t
s h o w n ) . I n t h i s c a s e , t h e r e f o r e , s o l u b l e p r o t e i n c o n t e n t
w a s n o t a r e l i a b l e f u n g a l g r o w t h i n d i c a t o r a s i t t e n d e dt o g i ve a n e r r o n e o u s r e s u l t. I t c o u l d b e u s e d a s a f u n g a l
g r o w t h i n d i c a t o r o n l y if t h e m e d i u m c o n t a i n e d a n i g n-
o r a b l e a m o u n t o f s o l u b l e p r o t e i n s . I n s t e a d , i t w a s m o r e
r e l i a b l e t o u s e t h e g l u c o s a m i n e c o n t e n t a s t h e g r o w t h
i n d i c a t o r . I t i s s h o w n i n F i g . 5 t h a t t h e a d d i t i o n o f
a b o u t 0 . 2 - 0 . 3 m l o f F P H p e r g r a m o f cr a n b e rr y
p o m a c e r e s u l t e d i n t h e o p t i m a l g r o w t h o f a l l s e l e c t e d
fung i .
The growth o f all tested fungi o n a n optimized cranberry
p o m a c e m e d i u m
W i t h t h e g o a l o f o b t a i n i n g a g e n e r a l c r a n b e r r y - b a s e d
m e d i u m f o r g r o w t h o f s e l e c t e d f u n g i , a n o p t i m i z e d
m e d i u m f o r m u l a w a s d e v e l o p e d a s d e s c r ib e d i n t h e
M a t e r i a l a n d m e t h o d s s e c t io n . T h e o p t i m i z a t i o n o f t h e
m e d i u m w a s b a s e d o n t h e h y p o t h e s i s t h a t a l l t h e f u n g i
s e l e c t e d h a d s i m i l a r g r o w t h p a t t e r n s i n r e s p o n s e t o
E 6g
E
o 2J . X ~
I0 20 30 40 50
FPH ml/100g pomace)
Fig. 5 . The effect of FPH addi t ion on glucosamine produc-tion by Trichoderma Rhizopus and Penicillium strains o n cran-berry pomace.
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 6/7
8/13/2019 Producción Hongos En Pomaza De Cranberry
http://slidepdf.com/reader/full/produccion-hongos-en-pomaza-de-cranberry 7/7
Z Zheng and K Shetty 329
8. Xavie r , S . an d Lon sane , B . K. , Sug ar -canep r e s s mu d a s a n o v e l a n d i n e x p e n s iv e s u b s t r a t e f o rproduc t ion of lac t ic ac id in a so l id s ta te f e rmenta -t ion sys tem. Appl Microbiol Biotechnol 1994, 41,291-295.
9 . Hang, Y. D. , Luh, B . S . and Woodams, E . E . ,
Mic robia l p roduc t ion of c i t r ic ac id by so l id s ta tefe rmenta t ion of k iwif ru i t pee l . J . F o o d S c i 1987,52, 226-227.
10 . Canne l , E . and Moo-Young, M. , Sol id s ta te f e r -me n ta t i o n s y s te ms . Proc Biochem 1980, 4 , 2 -7 .
11. Ngad i, M. O . and Co rreia , L. R. , Solid sta tee th a n o l f e r me n ta t i o n o f a p p l e p o ma c e a s a f f e c t e db y mo i s tu r e a n d b io r e a c to r mix in g s p e e d . J . F o o dSci 1992, 57, 667-6 70.
12. Worrall , J . J . and Yang, C. S. , Shii take and oysterm u s h r o o m p r o d u c t i o n o n a p p l e p o m a c e a n dsawdust . HortScience 1992, 27, 1131-1133.
13 . Hang, Y. D. and Woodams, E . E . , / F ruc tofur -a n o s id a s e p r o d u c t i o n b y Aspergillus s p e c i e s f r o ma p p le p o ma c e . Food Sc i Techno l 1 9 9 5 , 2 8 ,340-342.
14. Uj i ie , M. , Roy, C . and Y aguchi , M. , Low -mo lecula rwe ight xy lanase f rom Trichoderma viride ApplEnviron Microbiol 1991, 57, 1860-1862.
15 . Vas i leva -Tonkova , E . S . and Bezborodova , S . I . ,Pur i f ica tion , phys icochem ica l p rope r t ie s , and spec i -f ic i ty of a r ibonuc lease produced by Trichodermaharz ianum Enzy me Microb Tec hno l 1996, 18 ,147-152.
16 . Ulhoa , C . J . and Peberdy , J . F . , Pur i f ica t ion ands o me p r o p e r t i e s o f t h e e x t r a c e ll u l a r c h i t in a s e p r o -d u c e d b y Trichoderma harzianum Enz ym e Microb
Technol 1992, 14, 236-24 0.17 . Andrews, J . H . , B io logica l cont ro l in the phyl lo-
sphe re . An nu Rev Phytopathol 1992, 30, 603 -635.1 8 . K a t a y a ma , A . a n d Ma t s u mu r a , F . , D e g r a d a t i o n o f
organochlor ine pes t ic ides , pa r t icu la r ly endosul fan ,
by Trichoderma harzian um Environ Toxicol Chem1993, 12, 1059-10 65.
19. Soccol, C. R. , Marin, B. , Raimbault , M. and Leb-eaul t , J . M. , Breeding and growth of Rhizopus inraw cassava by so l id s ta te f e rmenta t ion . ApplMicrobiol Biotechnol 199 4, 41, 330 --336.
20 . Zheng, Z . , E legado, F . B . and Fuj io , Y. , P roduc-t i o n a n d s o me p r o p e r t i e s o f c e l l u l a s e f r o m R h i -zopus japon icus IFO5318. A n n u R e p IC B i ot e ch1993, 16, 223-2 32.
21 . Zheng, Z . , Levin , R . E . & She t ty , K. , Decolor iza -t ion o f po lym er ic dyes by a nove l Penicillium isola tein l iqu id media . P resented a t the Mic robia l Poly-me r s a n d G r a d u a t e E d u c a t i o n i n a G lo b a lE c o n o my , a s p e c i a l c o l l o q u iu m o r g a n i z e d b y t h eNew England Soc ie ty for Indus t r ia l Mic robio logy ,Ma y 2 9 , 1 9 97 , A m h e r s t , M A
22. Mar t in , A. M. and Chin ta lapa t i , S . P . , F ish of fa l -p e a t c o mp o s t e x t r a c t s a s f e r me n ta t i o n s u b s t r a t e .Biol Wastes 1989, 27, 281 -288.
23 . Bradford , M. M. , A rap id and sens i t ive me thod fort h e q u a n t i t a t i o n o f mic r o g r a m q u a n t i t ie s o f p r o t e inut il iz ing the pr inc ip le of pro t e in -dy e b inding . A n a lB iochem 1976, 72, 248-254 .
24. McCune, T. D. , Lang, K. W. and Steinberg, M. P. ,W a te r a c t i v i t y d e t e r min a t i o n w i th t h e p r o x imi tyequ il ibra tio n cell. J. Food Sc i 1981, 46, 1978-1979.
25 . Sakura i , Y . , Lee , T . H. and Shio ta , H . , On thec o n v e n i e n t me th o d f o r g lu c o s a min e e s t ima t io n i nKoji. Agric Biol Chem 1977, 41, 619 -624.
26 . Desgranges , C . , Vergoignan , C . , Georges , M. andDurand, A. , B iomass e s t ima t ion in so l id s ta te f e r -me n ta t i o n . Appl MicrobioL Biotechnol 1991 , 35 ,
200-205.27 . Roc he , N . , Venagu e , A. , Desgrang es , C . and
D u r a n d , A . , U s e o f ch i t in m e a s u r e me n t t o e s t ima t efunga l b iomass in so l id s ta te f e rmenta t ion . BiotechAdv 1993, 11, 677-683.