-------The 3rd ICMBMP

October 1999

Growth of Lentinus edodes on the coffee industry

residues and fruiting body production

Fan Leifa, Ashok Pandey and Carlos R. Soccol*

Laboratorio de Processos Biotecnologicos,

Departamento de Engenharia Quimica,
Universidade Federal do Parana, CEP 81531-970
Curitiba-PR, Brazil

* Corresponding author.Fax +55-41-266 0222

E-mail: soccol@engquim.ufpr.br
 Growth of Lentinus edodes on the coffee industry
residues and fruiting body production
Fan Leifa, Ashok Pandey and Carlos R. Soccol*
Laboratorio de Processos Biotecnologicos, Departamento de Engenharia Quimica,
Universidade Federal do Parana, CEP 81531-970 Curitiba-PR, Brazil

ABSTRACT

The strain of L. edodes LPB 02 was selected for fructification of these experiments. The
spawn for inoculation was made of sawdust (80%) and rice bran (20%). The substrates for
fructification were coffee husk (treated and untreated), coffee spent ground, a mixed
substrate comprising coffee husk (50%) and coffee spent ground (50%). The spawn rate of
10% was adopted; the humidity for mycelial growth was 55-65%. The cultivation was
carried out in plastic bags. The results showed that after 20 days growth on untreated coffee
husk, the mycelia regressed and humidity was augmenting. There was no fruiting body.
Treatment of coffee husk with hot water (cooked in water and filtered) was found useful for
its utilization as the substrate by the fungal culture. This resulted good mycelial growth and
good fruiting bodies were obtained. The biological efficiency reached at 85.8%. When
coffee spent ground was used as the substrate, the mycelia were relatively much white after
20 days of growth. First fructification occurred after 50 days of inoculation and second
after 70 days of inoculation. The biological efficiency reached at 88.6%. But when mixture
of spent ground and coffee husk was used as the substrate, the biological efficiency
achieved was less than that obtained with coffee spent ground (78.4%).

_________________________________________________________________________
*Corresponding author. E-mail: soccol@engquim.ufpr.br, Fax +55-41-266 0222
 INTRODUCTION

Coffee husk, which is the major residue of the coffee industry, is one of the most important
residues in more than 50 coffee-producing countries, and is generated about 1 million ton
yearly (Tango, 1971; ICO, 1998). Due to its toxic components, such as caffeine, tannins
and polyphenols, coffee husk does not find any application and has practically been not
utilized for any purpose (Soccol, 1995. Its disposal rather causes environmental pollution in
nature. Coffee spent ground, another residue of the coffee industry, is almost in the same
quantities as coffee husk, distributed worldwide, which also is of equal concern for
environmental pollution (Tango, 1971; Soccol, 1995; ICO, 1998). Brazil is a country
producing the largest quantity of coffee spent ground in the world (ICO, 1998). This
material was often composted as a soil conditioner, but it is also given to the waste depots,
resulting in lost of organic material (Thielke, 1989).

Lentinus edodes is one of the most important edible mushrooms cultivated world-wide
(Yang, 1985; Zhang 1989; Fan, 1990). It has excellent organoleptical properties of flavour
and aroma, and possesses good nutritional and therapeutically values due to the presence of
all essential amino acids required for humans. It also shows antitumoral activity, helps in
reducing the cholesterol and strengthening the immunological system (Zhang 1989). L.
edodes has been traditionally cultivated in the trunks of some hard trees (Yang, 1985). But
a lot of reports have appeared on its cultivation on some agricultural residues in the world,
especially in the mainland China (Yang, 1985; Zhang 1989; Beaux, 1994, 1996).

Mata (1994) has obtained the fruit body using coffee pulp which comes from wet
processing in the coffee industry. During the wet processing a lot of liquid residue was
produced, causing environmental pollution. In Brazil, the coffee husk comes from the dry
method, full of the nutrients, also full with caffeine, tannins, polyphenols, which is very
different in the compositions from that of other countries (Vasco, 1989. The extracted
coffee grounds, moistened with solution of 0,5% yeast extract at a ratio of 1:2 (W/V), were
used for culture of L.edodes and the fruiting bodies were obtained (Thielke, 1989). But this
technique is more expensive and not practical for production due to the addition of yeast
extract. This work was to explore some techniques more practical for its cultivation in the
Brazilian conditions or for the other developing countries and check possible biological
efficiency. Water content and spawning rate are the parameters most important in the
cultivation of edible mushroom, which affects directly the time of colonisation of mycelia
(vegetative phase ) and initiation of fructification. This work objects to determine those
factors for better application of those residues.

MATERIALS AND METHODS

Micro-organism L.edodes LPB 02, routinely maintained at 4 oC on PDA, was selected for
this study because of its good growth characteristics on the extract of coffee husk.

Spawn preparation The sawdust of Eucalyptus sp ( 80%) and rice bran ( 20% ) were used
for the spawn preparation. The mixture was adjusted at the moisture of 60% (Yang, 1985 )
and then filled in the flask. After autoclave and inoculation, the flask were incubated in the
dark at 24 oC. The spawn in the flask was ready for inoculation to objective substrate after
20 days growth when the mixture has turned white totally.

Substrate preparation The spent ground came from the solvent coffee factory after sun-dry.
The effect of water content in the substrate and rate of inoculation ware conducted in the
experiments. The concentration of water is at 45, 50%, 55%, 60%, 65%, 70% and 75%
respectively. The rate of inoculation is at 2,5, 5%, 10%, 15%, 20% and 25% respectively.
The substrate was moistened and 4 hours later, was filled in autoclavable plastic bag (Cai,
1992). The bags were autoclaved at 121 oC for 1,5 hours. When inoculation, the spawn was
separated into smallest particles. After inoculation, good mixture between spawn and
substrate was made so that mycelial growing could be evaluated. After inoculation, the
bags were incubated in the dark at 24 oC. mycelial development in the bag was observed
each day after 5 days.

Fructification The 100 g of dry spent ground each bag was used for the fructification. The
humidity and spawn rate were adopted according to the results gained above. At the 30
days, the bags were transferred to a lighted environmental chamber, cotton plugs loosened
to allow better aeration. After total transformation from whiteness to brown colour, the
plastic was removed for fructification.

RESULTS

GROWTH AND FRUCTIFICATION OF L. edodes LPB 02 ON THE COFFEE HUSK

When coffee husk was used as the substrate, the ideal humidity for mycelial growth was
worked out at the 55-65%; the best spawn rate at 10-15% was more efficient while it took
20 days for full occupation of mycelia; with the time of growing, the mycelia regresses and
secrets water so no fruit body has been obtained.

When coffee husk was proceeded treatment by boiling one hour in the water, then filtrated
and the solid residue was used for cultivation of this fungus. The ideal humidity for
mycelial growth was worked out at the 65-75%; the best spawn rate at 10-20% was more
efficient while it took 20 days for full occupation of mycelia; then the transformation of
colour was beginning. first fructification occurred after 60 days of inoculation; the
biological efficiency reached at the 85,76%.

Figure 1 Fructification of L. edodes LPB 02 on the treated coffee husk

GROWTH AND FRUCTIFICATION OF L. edodes LPB 02 ON THE COFFEE SPENT
GROUND

When coffee spent ground was used as the substrate, the ideal humidity for mycelial growth
was worked out at the 55-65%; the best spawn rate at 10% was more efficient and
economic while it took 20 days for full occupation of mycelia; first fructification occurred
after 56 days of inoculation; the biological efficiency reached at the 88,64%.

Figure 2 Fructification of L. edodes LPB 02 on the coffee spent ground

GROWTH AND FRUCTIFICATION OF L. edodes LPB 02 ON THE MIXTURE OF

COFFEE SPENT GROUND (50%) AND COFFEE HUSK (50%)

When coffee husk mixed with coffee spent ground was used as the substrate, the ideal
humidity for mycelial growth was worked out at the 55-65%; the best spawn rate at 10-20%
was more efficient and economic while it took 20 days for full occupation of mycelia; first
fructification occurred after 65 days of inoculation; the biological efficiency reached at the
78,36%.
Figure 3 Fructification of L. edodes LPB 02 on the mixture of coffee spent ground (60%)
and coffee spent ground (40%)

DISCUSSION AND CONCLUSIONS

The studies above showed that treated coffee husk and spent ground are good substrates for
the cultivation of this fungus. Those materials are plentiful in Brazil and available all year
round. The techniques for the cultivation is very simple, inexpensive, easily extended to the
other region, such as Africa and Central America. The untreated coffee husk appeared not
as good as treated one maybe because of much more content of caffeine, tannins and
polyfenols. Various testes have been made but no fruit body was obtained.

According to observation of growth of mycelia of this fungus, the spent ground also can be
used as the material of the spawn for inoculation. To achieve more production, maybe
additional wheat bran would have a more positive effect. For commercial purpose it must
be more effective to replace the laboratory inoculation by a special spawning way or
develop a mechanic way. Also more large bag should be used.

The results showed that coffee husk and coffee spent ground can be utilized for the
cultivation of this valuable mushroom. This offers an interesting opportunity to utilize it in
economical way.
 ACKNOWLEDGMENTS

Financial assistance from the European Union (grant no INCO DC: IC18*CT 970185) is
gratefully acknowledged. CRS thanks CNPq for a scholarship under Scientific Productivity
scheme.

REFERENCES

BEAUX, M.R. ; SOCCOL, C.R. ; RAIMBAULT, M. Use cassava bagasse on cultivation

edible fungus Lentinus edodes by solid state fermentation. In : G.M. ZANIN & F.F.
MORÌES (Eds). Anais IV Semin‡rio de Hidr—lise Enzim‡tica de Biomassas. Editora
Sthampa, 1994, p 190-198.

BEAUX, M. R. & SOCCOL, C. R. - Cultivo do fungo comest’vel Lentinula edodes em

res’duos agroindustriais do Paran‡ atravŽs do uso de fermenta•‹o no estado s—lido. Boletim
do CEPPA, vol. 14 (1) p. 11-21,1996.

Cai, W. M. and Fang, J.L. and Fan, L.F 1992 The disease of soft rotting in Flammulina
Velutipes Edible mushroom, Shanghai, China.12:129-132.

Chang, S.T. 1989 Edible mushroom and their cultivation CRC Press, Florida. 345p.

Fan, L.F. and Ding, C.K 1990 Handbook of mushroom cultivation Jiangxi science and
technology publishing house, Jiangxi, China. 388p.

International Coffee Organization 1998 Total production of exporting members

http://www.ico.org/proddoc.htm
Mata, G. , Gaitan Hernandez R. 1994 Advances in shiitake cultivation on coffee pulp
Revista Iberoamericana de Micrologia, 11: 90-91.

SOCCOL, C. R. Aplicaciones agroindustriales de los processos de fermentation em medios

solidos. In: Fronteras en Biotecnologia Y Bioengeneria. Sociedade Mexicana de
Biotecnologia y Bioengeneria, Mexico, 1995. p126-141.

SOCCOL, C. R. Aplica•›es da fermenta•‹o no estado s—lido na valoriza•‹o de res’duos

agroindustriais. Fran•a -Flash Agricultura, 4(2)Ê: 3-4, 1995.

Tango, J. S. 1971 Utiliza•‹o industrial do cafŽ e dos seus subprodutos Boletim do Instituto
de Tecnologia de Alimentos-ITAL S‹o Paulo, Brasil. 28:49-73.

Thiclke, C. Cultivation of edible fungi on coffee grounds Mushroom Science XII (parte II)
Germany. 12 (1989), 337-343

VASCO, J.Z. Utiliza•ion Integral de Los Sub-produtos do CafŽ . I Semin‡rio Internacional

Sobre Biotecnologia en la Agroindustria Cafetalera. Xalapa, MŽxico, 12 a 15/04/1989,
p.63-76.

Yang, X. M. 1986 Cultivation of edible mushroom in China Agriculture printing house,

Beijing, China.