World J Microbiol Biotechnol (2007) 23:14091415

DOI 10.1007/s11274-007-9381-z

ORIGINAL PAPER

Fermentation of cashew apple juice to produce high added value

products
Talita Lopes Honorato Maria Cristiane Rabelo
Luciana Rocha Barros Goncalves Gustavo Adolfo Saavedra Pinto
Sueli Rodrigues

Received: 4 October 2006 / Accepted: 21 December 2006 / Published online: 7 April 2007

Springer Science+Business Media B.V. 2007

Abstract The use of agriculture substrates in industrial

biotechnological processes has been increasing because of
its low cost. Cashew apples are considered an agriculture
low cost product in the Brazilian Northeast because the
cashew cultivation is done mainly to produce cashew nuts.
About 90% of the cashew apples production is lost in the
field after removing the nut. In this work, the use of clarified cashew apple juice as substrate for microbial cultivation was investigated. The results showed that cashew
apple juice is a good source of reducing sugars and can be
used to grow Leuconostoc mesenteroides to produce high
added value products such as dextran, lactic acid, mannitol
and oligosaccharides.
Keywords Low cost substrate
Fermentation

Response surface analysis
Cashew apple juice

High added value products

Introduction
Alternative substrates have been studied as low cost substrates for microbial fermentation to produce several
T. L. Honorato
M. C. Rabelo
S. Rodrigues (&)
Departamento de Tecnologia de Alimentos, Universidade
Federal do Ceara, Caixa Postal 12168, Fortaleza 60021-970,
Brazil
e-mail: sueli@ufc.br
L. R. B. Goncalves
Departamento de Engenharia Quimica, Av. Mister Hull, 2977,
bloco 709 Campus do Pici, Fortaleza, CE 60455-760, Brazil
G. A. S. Pinto
Embrapa Agroindustria Tropical, Rua Sara Mesquita,
2270 Pici, Fortaleza 60511-110, Brazil

products for the chemical, petrochemical, pharmaceutical

and food industry (Angumeenal and Venkappayya 2005;
Adham 2004; Kuerbanoglu 2004; Hang and Woodams
2000; Kumasr et al. 2003; El-Samragy et al. 1996; Stredanska et al. 1993; Vazquez et al. 2006; Tony et al. 2004;
Stredansky et al. 1999). Agriculture residues and wastes
are the most suitable low cost substrates for microbial
cultivation, being a source of carbon, nitrogen and minerals. However, despite the virtual potential use of any
agriculture resource as low cost substrate, their complex
composition may have some substances that may interfere
with the fermentation inhibiting microbial growth inactivating the enzymes associated with product synthesis.
Thus, some strains are able to grow in some agriculture
substrates and might not be able to grow in other. Besides,
the microbial growth can occur without the production of
the desired metabolite. The study of the use of agriculture
substrates to grow a particular strain of microorganism is
an important step before considering the industrial application of the raw material.
Cashew apple is a pseudo-fruit rich in reducing sugars
(fructose and glucose), vitamins, minerals and some amino
acids (Campos et al. 2002). The cashew tree grows into
quite a large area even on poor soils with low rainfall and is
cultivated in 32 countries around the world being Brazil,
India, Vietnam and Nigeria the main producers. Although
cashew apples can be consumed as juice, ice cream and
other foodstuffs, the cashew tree cultivation is an agricultural activity that aims mainly the production of cashew
nuts. The nuts represent only 10% of the total fruit weight,
and large amounts of cashew apples are left in the field
after nut removal.
According to official data, the Brazilian Northeast
presents an annual production of about two millions of tons
of cashew apples and 90% of this production is lost or

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underutilized. As such, cashew apples are considered an

agriculture residue and, its nutritive juice can be a suitable
low cost substrate for microorganisms growth.
Glycosyltransferases are enzymes that catalyze the
transfer of glycosyl residues from a donor molecule to a
particular acceptor (Rabelo et al. 2006; Rodrigues et al.
2005, 2006; Monchois et al. 1999; Monsan and Paul 1995;
Koepsell et al. 1953). Lactic acid bacteria produce a wide
variety of a particular group of glycosyltransferases, which
can synthesize oligosaccharides and polymers. Leuconostoc mesenteroides B512F is a lactic acid bacterium that
produces dextransucrase (EC 2.4.1.5), dextran, fructose
and prebiotic carbohydrates. This bacterium also reduces
fructose to mannitol during the fermentation. Mannitol is a
six carbon polyol with several applications in the food and
pharmaceutical industry. Mannitol is a valuable nutritive
sweetener because it is non-toxic, non-hygroscopic in its
crystalline form and has no teeth decaying effects. It has a
sweet, cool taste and it is about half as sweet as sucrose.
Mannitol is applied as a food additive, as a sweet tasting
bodying and texturing agent and is used as a sweet builder
in sugar free chewing gum and in pharmaceutical
preparations (Wisselink et al. 2002).
As the industrial strain employed to produce dextran and
dextransucrase is Leuconostoc mesenteroides NRRL
B512F, this strain was employed to evaluate the potential
use of cashew apple juice as a fermentative substrate for
producing high added value compounds for food industry
as prebiotic-oligosaccharides, lactic acid, dextran and
mannitol. The viability of using this alternative and low
cost material was evaluated through a factorial planning
where sucrose was added to the medium to induce the
production of the dextransucrase enzyme (Pereira et al.
1998; Rodrigues et al. 2003). The effect of phosphate
(K2HPO4); nitrogen (yeast extract) and the addition of
minerals were also evaluated.
Materials and methods

World J Microbiol Biotechnol (2007) 23:14091415

with deionized water followed by immersing the glasses in

a deionized water bath containing HCl 10%. The glasses
were allowed to stand 24 h in this solution and then were
rinsed five times with deionized water.
The samples were previously mineralized digesting
5 mL of the cashew apple juice with 8 mL of a digesting
solution composed of HNO3: HClO4 (3:1 v/v). This procedure was carried out in a hood using a digester dry block
(Tecnal model TE007D) at 200 C until the final volume
reached 1.5 mL (34 h). This volume was quantitatively
transferred to a 50 mL volumetric flask and the final volume was completed with deionized water. A blank sample
was prepared in the same way, using deionized water instead of the juice.
Fe, Ca, Mg, Mn, Zn and Cu were quantified in an atomic
absorption spectrometer (Perkin-Elmer, model A-Analyst
300). Fe, Cu and Zn were directly determined from the
diluted samples prepared as described above. Ca and Mg
were previously diluted in water followed by a second
dilution in a 1.14 g/L lantan solution.
Potassium and sodium were determined by flame photometry in a DIGIMED model DM-61 equipment. The
digested samples prepared as described above were previously diluted in deionized water.
Determination of nitrogen content of the clarified
cashew apple juice
The nitrogen content of the clarified cashew apple juice
was determined according to the Kjeldahl method (AOAC
1995). Results were expressed as total protein (f = 5.75).
Determination of pH and reducing sugar of clarified
cashew apple juice
Total reducing sugar was measured by the DNS method
(Miller 1959) using a UV-visible spectrophotometer
(Micronal B-542). The pH was determined by direct
measure in a Marconi PA 200 potentiometer.

Cashew apple juice

Fermentative assays
The cashew apple juice was obtained through mechanical
process. This juice, which contains high levels of tannins,
was clarified by adding gelatin to remove tannins and
suspended solids. The clarified cashew apple juice was
physico-chemically characterized; filtered and stored
frozen (20 C) prior to use.
Determination of the mineral content of the clarified
cashew apple juice
All the materials used to prepare the samples were previously washed with neutral detergent and rinsed five times

123

A strain of Leuconostoc mesenteroides B512F obtained

from ARS Culture Collection (NRRL Culture collection,
United States Department of Agriculture, Peoria, Illinois USA) was activated in an optimized synthetic medium
composed of sucrose, 50 g/L (food grade); yeast extract,
20 g/L (Himedia); MgSO4.7H2O, 0.20 g/L; MnSO4.2H2O,
0.01 g/L; FeSO4.7H2O, 0.01 g/L; CaCl2.2H2O, 0.02 g/L;
NaCl, 0.01 g/L; and K2HPO4 (anhydrous), 20 g/L (Rodrigue et al. 2003; Rodrigues et al. 2005; Rabelo et al. 2006)
in an orbital shaker at 32 C and 150 rpm. The strain is
viable for at least three consecutives shake cultures at the

World J Microbiol Biotechnol (2007) 23:14091415

1411

essay condition (Rodrigues et al. 2003). This cell culture

was used as inoculums to the cashew apple juice medium.
The inoculums volume was 10% of the total culture volume.
The fermentation medium using cashew apple juice in
substitution to the synthetic culture medium was prepared
diluting the clarified apple juice to reach the desired
reducing sugar content and adding sucrose, yeast extract,
phosphate and minerals.
The fermentative assays were carried out according to a
fractional factorial experimental design (2(52)) where five
parameters were changed in two levels. Total reducing
sugar (fructose + glucose), sucrose, yeast extract, phosphate (K2HPO4) and minerals were changed according to
the experimental design presented in Table 1. The levels of
the parameters were based on the synthetic medium composition (Rodrigues et al. 2003, 2005; Rabelo et al. 2006).
Sucrose was added to induce dextransucrase production,
allowing dextran and oligosaccharide formation. The initial
pH was adjusted to 6.7 (optimum pH for L mesenteroides
B512F growth) and the culture medium was sterilized at
121 C for 15 min. Phosphate solutions were sterilized and
added to the sterile fermentation medium after cooling to
room temperature.
Fermentation was carried out in an orbital shaker
(Tecnal TE-420) at 32 C and 150 rpm for 24 h in 250 mL
Erlenmeyrs flasks containing 100 mL of the culture medium. Biomass, dextran, oligosaccharide, mannitol and lactic acid production, as well as, residual sucrose, glucose
and fructose were the responses evaluated through response
surface analysis. The data were handled using the software
Statistica v 5.0 (Statsoft) considering a 95% confidence
level.
Biomass was determined by direct optical density at
660 nm. After harvesting the cells by centrifugation at
11806 g (8000 rpm) for 10 min, the supernatant was used
to determine the carbohydrates content. Lactic acid was
assayed by titration with NaOH.

Table 1 Experimental design

for clarified cashew apple juice
fermentation with L.
mesenteroides B512F

* Mineral salts as percentages

based on the synthetic medium

Carbohydrate analysis
After harvesting the cells, dextran was precipitated by
adding three volumes of ethanol (96% v/v). The pellet was
re-suspended in distilled water and the dextran formed
during the fermentation was assayed as total carbohydrate
(Dubois et al. 1956).
The supernatant containing oligosaccharides, fructose,
glucose, sucrose and mannitol was analyzed by high performance liquid chromatography (HPLC) with a Varian
ProStar system equipped with two high-pressure pumps
model 210, refraction index detector model 350 and a
column oven Eldex CH model 150. Separation was
achieved on an AminexHPX 87 C (300 mm 7.8 mm)
column at 85 C. Ultra pure water at 0.3 ml/min was used
as eluent and the detector temperature was 45 C. The
software ProStar WS 5.5 was used to acquire and handle
the data. The initial amounts of glucose and fructose in the
cashew apple juice were also assayed by HPLC.

Results and discussions

High performance liquid chromatography (HPLC) showed
that the cashew apple juice contained equal amounts of
glucose and fructose (reducing sugars). Table 2 presents
the nutritional composition of the clarified cashew apple
juice used in this work. The mineral content of the cashew
apple juice presented higher levels of magnesium and sodium than the synthetic medium. On the other hand, the
levels of iron, manganese, potassium and phosphorous
were lower in the cashew apple juice than in the synthetic
medium. Calcium was not detected in the cashew apple
juice. Zinc was not present in the synthetic medium but
was found in the cashew apple juice. The nitrogen content
was lower in cashew apple juice than in synthetic medium.
Considering the juice dilution, except for sodium and
magnesium, the medium formulated with cashew apple

Run

Sucrose (g/L)

Reducing sugar (g/L)

Yeast extract (g/L)

Phophate (g/L)

Salt* (%)

50.0

50.0

20.0

20.0

100

50.0

50.0

0.0

20.0

50.0

25.0

20.0

0.0

100

50.0

25.0

0.0

0.0

25.0

50.0

20.0

0.0

25.0

50.0

0.0

0.0

100

7
8

25.0
25.0

25.0
25.0

20.0
0.0

20.0
20.0

0
100

37.5

37.5

10.0

10.0

50

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World J Microbiol Biotechnol (2007) 23:14091415

Table 2 Nutrient composition of the clarified cashew juice used to

formulate the cashew culture medium
Macronutrients
Component

Phosphorous
Potassium

Micronutrients
Content
(g/L)

Component Content

1.21 0.01 Cooper

13.13 0.87 Ferrous

<DL
6.97 2.68 103

Calcium
Magnesium

<DL
Zinc
11.20 4.31
1.17 0.07 Manganese 6.40 0.35

Sodium

0.09 0.0

Sulfur

0.81 0.02

Reducing sugar

90.45 5.25

Nitrogen as total
protein

2.58 0.58

juice without the addition of external source of phosphorous, nitrogen and minerals was significantly poor compared to the synthetic medium.
Table 3 presents the results obtained for each run carried
out according to the experimental planning (Table 1).
According to them, the fermented broth presented biomass
growth; dextran and oligosaccharide synthesis, besides
lactic acid and mannitol production. None of the initial
substrates were totally consumed.
Higher biomass values were obtained when equal
amounts of reducing sugars and sucrose were employed
and the medium was supplemented with yeast extract and
phosphate (Runs 1 and 7). Higher levels of oligosaccharides were also found in this experimental condition.
However, considering the amount of the initial carbohydrates, the yield of run 7 (14.82%) was higher than the
yield of run 1 (9.30%). Higher yields of lactic acid were
found in run 7 (30.54%) when compared to run 1 (20.75%).
Dextran yield, based only on the initial sucrose content,
was 8.64% in run 1 and 11.96% in run 7. Considering only
the initial reducing sugar content, oligosaccharide yields of
18.60% and 29.64% were found respectively in runs 1 and
7. The addition of external nitrogen source improved the
oligosaccharide yield. Comparing the oligosaccharide
Table 3 Dextran,
oligosaccharides, biomass and
lactic acid in the cashew apple
juice fermented broth

123

yields based on the initial amount of reducing sugar of runs

1 and 2 (18.60% and 9.20% respectively) to the yields of
runs 7 and 8 (29.60% and 20.80% respectively) it is clear
that this improvement was more effective (about 50%)
when high levels of carbohydrates (runs 1 and 2) were
present in the medium. Higher yields of oligosaccharides
were obtained when low carbohydrates were initially
present in the culture medium (runs 7 and 8). High levels of
mannitol (above 10 g/L) were found when high initial
levels of reducing sugars were used (runs 1,2, 5 and 9) or
high amounts of dextran were produced (run 7), since
fructose is a side product of dextran synthesis. Values of
pH below 4.0 were obtained for the runs without addition
of phosphate (runs 36) or when the phosphate concentration was lower than the observed in the synthetic medium (run 9).
The effect of each independent variable on the responses
presented in Table 3, except for pH values, is presented in
Table 4. The addition of mineral salts to the clarified
cashew apple juice culture broth was not significant at the
considered confidence level (95%) for any response,
meaning that the amount of minerals present in the juice
(Table 2), was sufficient to supply the mineral requirements of L. mesenteroides B512F. All other effects, except
the mean effect, were significant for dextran synthesis. On
oligosaccharide synthesis and biomass growth only the
mean effect; yeast extract and phosphate were significant.
On mannitol production only sucrose and mineral salts
were not significant and for lactic acid formation only
mineral salts and reducing sugar were not significant.
The highest individual effect on dextran and oligosaccharides synthesis was yeast extract. The clarified cashew
apple juice presented a low level of protein (Table 2) when
compared to the synthetic medium, thus the addition of a
nitrogen source, as yeast extract, enhanced the biomass
growth and consequently the dextransucrase (the enzyme
responsible for both dextran and oligosaccharide synthesis)
production. However, it was possible to obtain both dextran
and oligosaccharides without any addition of extra nitrogen
source as shown in runs 2, 4, 6 and 8 (Table 3).

Run Dextran (g/L) Oligos (g/L) Mannitol (g/L) Biomass (ABS 660 nm) Lactic acid (g/L) FinalpH
1

4.37 0.04

9.30 0.60

17.44 0.42

2.58 0.04

20.75 0.17

4.02

2
3

2.76 0.09
2.72 0.10

4.60 0.43
6.07 0.43

11.22 0.53
9.56 0.36

2.15 0.02
2.06 0.01

15.23 0.10
11.81 0.10

4.11
3.79

1.51 0.07

1.80 0.60

1.52 0.04

5.84 0.17

3.73

2.65 0.06

6.64 0.69

12.66 0.07

1.94 0.01

10.57 0.68

3.89

1.49 0.18

3.68 0.33

8.02 0.37

1.66 0.04

5.42 0.75

3.72

2.99 0.18

7.41 0.33

12.40 0.37

2.66 0.04

15.27 0.75

4.20

1.56 0.06

5.20 0.69

8.62 0.07

2.47 0.01

12.99 0.69

4.00

2.39 0.09

7.18 1.31

12.04 0.71

2.29 0.15

12.57 0.28

3.83

5.60 0.42

World J Microbiol Biotechnol (2007) 23:14091415

1413

The phosphate effect was the highest individual effect

on biomass growth and had an important effect on dextran,
oligosaccharide and mannitol production. This component
can be directly associated to the biomass growth since it
acts buffering the medium and keeping the pH near the
optimum value for L .mesenteroides B512F growth until a
satisfactory amount of biomass is obtained allowing the pH
to fall to the optimum enzyme activity (Rodrigues et al.
2003). The regression coefficients of the responses are
presented in Eqs. 15.
DXT  5:57  10

1

2:67  10

RS 6:75  10

2

2

S 2:48  10

2

YE 4:13  102

P 5:55  104 Salt

Table 4 Estimated effects of the studied variables on dextran, oligosaccharide, biomass and lactic acid production
Factor

Dextran

Oligosaccharides

Mannitol

5.7733*

10.8300*

Biomass

Lactic
acid

Effect
Mean

2.4903

Sucrose

0.6670*

0.2650

0.5300

0.1060

2.1460*

12.2827*
2.3740*

Reducing
sugar

0.6270*

0.9575

3.2900*

0.0960

1.5415

Yeast
extract

1.3505*

3.5125*

4.6500*

0.3622*

4.7130*

Phosphate

0.8255*

2.1025*

3.4600*

0.6675*

7.6800*

Salts

0.0555

0.9275

0.4400

0.1272

0.9900

* significant at 95% of confidence level

1
OLIG 1:46  1:06  102 S 3:83  102
RS 1:76  101 YE 1:05  101
P 9:28  10

3

Salt

MAN 0:83 2:14  102 S 1:32  101

RS 2:33  101 YE 1:73  101
P 4:43  10

3

Salt

BIOM 1:87  4:18  103 S  3:78  103

RS 1:81  102 YE 3:34  102
P 1:27  10

LAcid  2:67  10

1

1

RS 2:35  10

Salt

9:48  10 S 6:16  10
1

Fig. 1 Fitted surface response for dextran production (Eq.1) as a

function of initial phosphate and yeast extract concentration
(sucrose = 37.5 g/L; reducing sugar = 37.5 g/L)

2

YE 3:64  101

P 9:90  103 Salt

5
where S is the sucrose concentration (g/L), RS is the
reducing sugar concentration (g/L), YE is the yeast extract
concentration (g/L), P is the phosphate concentration (g/L)
and Salt is the mineral salt added (%).
In the presented equations Biomass (BIOM) is expressed
as optical density at 660 nm, dextran (DXT), oligosaccharides (OLIG), mannitol (MAN) and lactic acid (LAcid)
are expressed in g/L.
F-test was used as significance criteria for the fitted
models. The calculated F-values (F5,3) for Eqs. 15 were
58.28; 9.3; 17.43; 26.67 and 48.44 respectively. At 95% of
confidence level the listed F5,3 is 9.01. Thus all the
regression models were significant since the calculated F-

values are higher than listed F-value (Rodrigues et al.

2006). The correlation coefficients (R2) of Eqs. 15 were:
0.9895; 0.9396; 0.9669; 0.9779; 0.9878 respectively.
Considering each studied response, yeast extract and
phosphate addition presented the highest effect on all of
them (Table 4). Surface response graphs built using the
fitted models are presented in Figs. 1 5. High concentrations of phosphate and yeast extract enhanced dextran
formation, oligosaccharide formation, mannitol production,
biomass growth and lactic acid formation.

Conclusions
In this work the use of cashew apple juice as substrate for
growing L. mesenteroides NRRL B-512F was investigated.

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1414

Fig. 2 Fitted surface response for oligosaccharides production (Eq.

2) as a function of initial phosphate and yeast extract concentration
(sucrose = 37.5 g/L; reducing sugar = 37.5 g/L)

Fig. 3 Fitted surface response for mannitol production (Eq.3) as a

function of initial phosphate and yeast extract concentration
(sucrose = 37.5 g/L; reducing sugar = 37.5 g/L)

The effects of sucrose, reducing sugars (glucose + fructose), external nitrogen source (yeast extract), phosphate
(K2HPO4) and mineral salts on the production of dextran,
oligosaccharides, mannitol lactic acid and biomass growth,
in the fermentation of clarified cashew apple juice by L.
mesenteroides B512F were studied. The results clearly
showed that cashew apple juice can be employed as an
alternative low cost substrate for L. mesenteroides B512F
growth, in order to produce high-added value products such
as dextran, mannitol, lactic acid and oligosaccharides. The
addition of mineral salts did not present significant effect
on the evaluated responses. Thus the mineral content of the
clarified cashew apple juice can be considered sufficient to

123

World J Microbiol Biotechnol (2007) 23:14091415

Fig. 4 Fitted surface response for biomass growth (Eq. 4) as a

function of initial phosphate and yeast extract concentration
(sucrose = 37.5 g/L; reducing sugar = 37.5 g/L)

Fig. 5 Fitted surface response for lactic acid production (Eq. 5) as a

function of initial phosphate and yeast extract concentration
(sucrose = 37.5 g/L; reducing sugar = 37.5 g/L)

provide the microbial mineral requirements. The addition

of a nitrogen source and phosphate presented a significant
effect on the analyzed responses and has enhanced biomass
growth, dextran, oligosaccharides, mannitol and lactic acid
production.
The use of agriculture residues as substrates for biotechnological processes is increasing as an attempt to reduce agriculture waste and process costs. According to the
results presented in this work, the use of cashew apple juice
as a fermentation substrate for L. mesenteroides NRRL
B512F cultivation is viable and the synthesis of oligosaccharides, mannitol and lactic acid can be carried out using
this substrate.

World J Microbiol Biotechnol (2007) 23:14091415

Acknowledgments The authors thank FUNCAP (Fundacao Cearence de Apoio ao Desenvolvimento Cientfico e Tecnologico) for the
awarded scholarship, CNPq (Conselho Nacional de Pesquisa e Desenvolvimento Tecnologico) for the financial support, ARS culture
collection for providing the microorganism strain and Laboratorio de
Combustiveis e Lubrificantes (DEQ/UFC) for the use of the HPLC.

References
Adham NZ (2004) Attempts at improving citric acid fermentation by
Aspergillus niger in beet-molasses medium. Bioresour Technol
84:97100
Angumeenal AR, Venkappayya D (2005) Artrocarpus heterophyllus:
a potential substrate for citric acid biosyntehsies using Aspergillus niger. LWT Food Sci Technol 38:8993
AOAC International (1995) Official methods of analysis. AOAC
International, Arlington, VA
Campos DCP, Santos AS, Wolkoff DB, Matta VM, Cabral LMC,
Couri S (2002) Cashew apple juice stabilization by microfiltration. Desalination 148:6165
Dubois M, Gilles KA, Hamilton PA, Rebers PA, Smith F (1956)
Colorimetric method for determination of sugars and related
substances. Anal Chem 28:350356
El-Samragy YA, Khorshid MA, Foda MI, Shehata AE (1996) Effect
of fermentation conditions on the production of citric acid from
cheese whey by Aspergillus niger. Int J Food Microbiol 29:411
416
Hang YD, Woodams EE (2000) Corn husks: a potential substrate for
production of citric acid by Aspergillus niger. LWT Food Sci
Technol 33:520521
Koepsell HJ, Tsuchia HM, Hellman NN, Kasenko A, Hoffman CA,
Sharpe ES, Jackson RW (1953) Enzymatic synthesis of dextran:
acceptor specificity and chain initiation. J Biol Chem 200:793
801
Kuerbanoglu EB (2004) Enhancement of citric acid production with
ram horn hydrolysate by Aspergillus niger. Bioresour Biotechnol
92:97101
Kumasr D, Jain VK, Shanker G, Srivastava A (2003) Citric acid
production by solid state fermentation using sugar cane bagasse.
Process Biochem 38:17311738

1415
Miller GL (1959) Use of dinitrosalicilic acid reagent for determination of reducing sugar. Anal Chem 31:426428
Monchois V, Willemont RM, Monsan P (1999) Glucansucrases:
mechanism of action and structure function relationships. FEMS
Microbiol Rev 23:131151
Monsan P, Paul F (1995) Enzymatic synthesis of oligosaccharides.
FEMS Microbiol Rev 16:187192
Pereira AM, Costa FAA, Rodrigues MI, Maugeri F (1998) In vitro
synthesis of oligosaccharides by acceptor reaction of dextransucrase from Leuconostoc mesenteroides. Biotechnol Lett
20:397401
Rabelo MC, Honorato TL, Goncalves LRB, Pinto GAS, Rodrigues S
(2006) Enzymatic synthesis of prebiotic oligosaccharides. Appl
Biochem Biotechnol 133:3140
Rodrigues S, Lona LMF, Franco TT (2003) Effect of phosphate on
production of Leuconostoc mesenteroides NRRL B512F. Bioprocess Biosyst Eng 23:5762
Rodrigues S, Lona LMF, Franco TT (2005) The effect of maltose on
dextran yield and molecular weight distribution. Bioprocess
Biosyst Eng 28:914
Rodrigues S, Lona LMF, Franco TT (2006) Optimization of panose
production by enzymatic synthesis using neural networks. J Food
Eng 75:433440
Stredanska S, Slugen D, Stredansky M, Grego J (1993) Arachidonic
acid production by Mortierella alpina grown on solid substrates.
World J Microbiol Biotechnol 9:511513
Stredansky M, Conti E, Navarini L, Bertocchi C (1999) Production of
bacterial exopolysaccharides by solid substrate fermentation.
Process Biochem 34:1116
Tony WY, Fu XY, Lee SM, Yu J, Wei DZ (2004) Purification of L
(+) lactic aid from fermentation broth with paper sludge as
cellulosic feedstock using weak anion exchange Amberlite IRA92. Biochem Eng J 18:8996
Vazquez JA, Gonzalez MP, Murado MA (2006) Preliminary testes on
nisin and pediocin production using waste protein sources
factorial and kinetic studies. Bioresour Technol 97:605613
Wisselink HW, Weusthuis RA, Eggink G, Hugenholtz J, Gorbben JH
(2002) Mannitol production by acid lactic bacteria: a review. Int
Dairy J 12:151161

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