Biotechnology Letters Vol 7 No 4 253-254 (1985)

GRAPE POMACE: A NOVEL SUBSTRATE FOR MICROBIAL PRODUCTIONOF CITRIC ACID

Y. D. Hang* and E. E. Woodams

Department of Food Science & Technology
Cornell U n i v e r s i t y , Geneva, New York 14456

SUMMARY
Grape pomace was used as a substrata for microbial production of
c i t r i c acld. Of the f l v e cultures examined, AsoergIllus niger NRRL 567 was
found ?o produce the grea%est amount of c i t r i c acld from grape pomace In
the presence of methanol at a concentration of 3% ( v o l / w t ) . The y i e l d was
60~ based on the amount of fermentable sugar consumed.

INTRODUCTION
Grape pomace Is the residue l e f t from Juice e x t r a c t i o n and c o n s t l t u t e s
about 16% of the o r i g i n a l f r u l t (Rlce, 1976). In 1984, approximately 4,9
m i l l i o n metric tons of grapes are produced In the United States. Grape
pomace ls rich In carbohydrates, but i t s nitrogen and phosphorus contents
are low (Rice, 1976). At present, most pomace ls dumped desplte the In-
creasing disposal problem posed and e f f o r t s made at by-product u t i l i z a t i o n
(FamuyIma and Ough, 1982).

C i t r i c acid Is a commercially valuable product and has been produced

primarily by AspergIllus niger grown on a sucrose or molasses medium
(Kapoor et a l , 1982). The objective of t h l s study was to determine the
s u l t a b I I I % / of grape pomace as a substrate for c i t r i c acld production.

MATERIALS AND METHODS

Substrate Grape pomace used in t h i s work was obtained from our p i l o t plant
juice processing u n l t and stored a t -25~ u n t i l needed. Before I t s use,
the pomace was ground through the 0.635 cm screen of a FItz M i l l , Model D
(W. J. FI?zpatrlck Co., Chicago, I l l i n o i s ) . I t contained 17.3% fermentable
sugar as glucose and had a moisture content of 65.4$. The pH was 3.8.
Cultures Five c i t r i c acld producing strains of AsDergIIIus niger were
obtained form Dr. C. W. Hesseltlne, Northern Regional Research Center,
Peoria, I l l i n o i s . Each c u l t u r e was grown on a potato dextrose agar s l a n t
a t )O~ for 7 days. A spore Inoculum was prepared by adding 3 ml of
s t e r i l i z e d d i s t i l l e d water to each s l a n t and shaking for one minute.
Solid state fermentation Portions of 40 g of grape pomace were Introduced
Into 500 ml Erlenmeyer flasks and s t e r i l i z e d a t 121~ for 15 minutes. Each
flask was Inoculated wlth 0.2 ml of an appropriate spore Inoculum for 4
days. Methanol was added t o the flasks before fermentation. At the end of
the fermentation, the fermented materials were extracted wlth water and the
extracts were analyzed for residual sugar and c i t r i c acld.
Analvtical methods Sugar was determined as glucose by the phenol-sulfurlc
acld methanol (Dubols e? a l , 1956). C i t r i c acld was measured by ?he
colorlmetrlc method of Taussky (1949).

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RESULTS AND DISCUSSION
Table 1 shows the production of c i t r i c acid from grape pomace by f i v e
s t r a i n s of ~9~z~JJ_LJ~ niger. The y l e l d s of c i t r i c acid varied con-
slderably and depended on (1) the amount of methanol present in the
subs?rate, and (2) the s t r a i n of A. niger used. Pomace samples fermented
In the presence of methanol contained a much greater amount of c l t r l c acid
than those fermented without the a d d i t i o n of methanol. The e f f e c t of
methanol In increasing c i t r i c acid y i e l d s appears to be a general
phenomenon in s t r a i n s of A. nlaer and the use of methanol has become a
common practice In c l t T I c acid production ~Kapoor 9_t a l , 1982). Of the
f i v e cultures examined, A. niger NRRL 557 produced the greatest amount of
c l t T l c acid from grape pomace In the presence of methanol a t a concentra-
t i o n of 3~ ( v o l / w t ) . The y i e l d of c i t r i c acid was 60~ based on the amount
of fermentable sugar consumed. Lakshminarayana et al (1975) reported t h a t
the y i e l d of c i t r i c acid from a sugarcane molasses medium under s o l i d state
conditions in the presence of methanol was 58.7~.

Dubols, M., Giles, K. A. Hamilton, J. K., Roberts, D. A. and Smith, F.

1956. Anal. Chem. 28, 350-356.

Famuylma, O. and Ough, C. S. 1982. Am. J. Enol. V l t l c . 33, 44-46.

Kapoor, K. K., Chaudhary, K. and Tauro, P. 1982. C l t r l c acid. In:

Prescott and DunnWs Industrial Mlcrobloloov. v .
4th ed. G. Reed (ed).
pp. 709-747. Connecticut= AVl.

Lakshmlnarayana, K. Chaudhary, K., E t h l r a j , S. and Tauro, P. 1975.

Biotechnol. Bioeng. 17, 291-293o

Rice, A. C. 1976. Am. J. Enol. Y l t t c . 27, 21-26.

Taussky, H. H. 1949, J. B i o l . Chem. 181, 195-198.

Table 1. C i t r i c acld production from grape pomace by Asoerglllu~ niger

( I n i t i a l sugar content 173.4 g/kg)

Methanol Resi dual Yleld of

Straln % sugar g/kg C i t r i c acid
g/kg sugar consumed

NRRL 2001 0 15.7 11 2

3 50.g 413
NRRL 2270 0 14.4 150
5 23.6 511
NRRL 599 0 21 .g 304
5 60.0 498
NRRL 328 0 14.5 170
3 30.0 52.3
NRRL 567 0 19.1 275
3 1g.8 600

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