Yacon Flour and Corn Steep Liquor as Substrate for Inulinase and Biomass Production by Kluyveromyces Marxianus NRRL Y-7571 (original) (raw)
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Inulinase Production by Kluyveromyces marxianus NRRL Y-7571 Using Solid State Fermentation
Applied Biochemistry and Biotechnology, 2006
Inulinase is an enzyme relevant to fructose production by enzymatic hydrolysis of inulin. This enzyme is also applied in the production of fructooligosaccharides that may be used as a new food functional ingredient. Commercial inulinase is currently obtained using inulin as substrate, which is a relatively expensive raw material. In Brazil, the production of this enzyme using residues of sugarcane and corn industry (sugarcane bagasse, molasses, and corn steep liquor) is economically attractive, owing to the high amount and low cost of such residues. In this context, the aim of this work was the assessment of inulinase production by solid state fermentation using by Kluyveromyces marxianus NRRL Y-7571. The solid medium consisted of sugar cane bagasse supplemented with molasses and corn steep liquor. The production of inulinase was carried out using experimental design technique. The effect of temperature, moisture, and supplements content were investigated. The enzymatic activity reached a maximum of 445 units of inulinase per gram of dry substrate.
Inulinase production by Kluyveromyces marxianus NRRL Y-7571 using solid state fermentation
Applied Biochemistry and Biotechnology, 2006
Inulinase is an enzyme relevant to fructose production by enzymatic hydrolysis of inulin. This enzyme is also applied in the production of fructo-oligosaccharides that may be used as a new food functional ingredient. Commercial inulinase is currently obtained using inulin as substrate, which is a relatively expensive raw material. In Brazil, the production of this enzyme using residues of sugarcane and corn industry (sugarcane bagasse, molasses, and corn steep liquor) is economically attractive, owing to the high amount and low cost of such residues. In this context, the aim of this work was the assessment of inulinase production by solid state fermentation using by Kluyveromyces marxianus NRRL Y-7571. The solid medium consisted of sugar cane bagasse supplemented with molasses and corn steep liquor. The production of inulinase was carried out using experimental design technique. The effect of temperature, moisture, and supplements content were investigated. The enzymatic activity reached a maximum of 445 units of inulinase per gram of dry substrate.
Optimisation of inulinase production by Kluyveromyces bulgaricus
Web Science, 2002
Present work is based on observation of effects of pH and temperature of fermentation on the production of microbial enzyme inulinase by Kluyveromyces bulgaricus (former Kluyveromyces marxianus). Inulinase hydrolyses inulin, an oligosaccharide which can be isolated from plants such as Jerusalem artichoke, chicory or dahlia, into pure fructose (1). Fructooligosaccharides have great potential in food industry because they can be used as calorie-reduced and noncariogenic sweeteners. Fructose formation from inulin is a single step enzymatic reaction and yields are up to 95 % fructose. On contrary, conventional fructose production from starch needs at least three enzymatic steps, yielding only 45 % fructose (2). Process of inulinase production was optimised by using experimental design method. pH value of the cultivation medium showed to be the most significant variable and it should be maintained at optimum value, 3.6. The effect of temperature was slightly lower and optimal values are between 30 and 33 ºC. At a low pH value of the cultivation medium, the microorganism was not able to produce enough enzyme and enzyme activities were low. Similar effect was caused by high temperature. Highest values of enzyme activities were achieved at optimal fermentation conditions and the values were: 100.16-124.36 IU/ml (with sucrose as substrate for determination of enzyme activity) or 8.6-11.6 IU/ml (with inulin as substrate), respectively. The method of factorial design and response surface analysis makes it possible to study several factors simultaneously, to quantify the individual effect of each factor and to investigate their possible interactions (3). The model based on physiological assumptions is also applied. Assumed is a single enzyme rate determing growth (Monod kinetics) with proportional inulinase production rate. Applied are the models of reversible temperature and acidity inhibition based on thermodynamic equilibrium between active and inhibited enzyme states. Predictions by the two models are compared by ANOVA.
Enzyme and Microbial Technology, 2006
The production of enzymes by bioprocesses is a good alternative to add value to agroindustry residues. Sugarcane bagasse is an abundant by-product of sugar industry and was tested as support and carbon source for production of inulinase (2,1--d-fructanohydrolase, E.C. 3.2.1.7) from Kluyveromyces marxianus NRRL Y-7571 by solid-state fermentation. Corn steep liquor was used as nitrogen supplement. Factorial design and response surface analysis were carried out to evaluate the effects of temperature (30.4-41.6 • C) and corn steep liquor (13-27.1%, w/v) on the production of inulinase. Optimum fermentation conditions were found to be: 36 • C and 20 wt.% of corn steep liquor. Under optimized conditions, the extra-cellular enzyme concentration reached 391.9 U/g of dry fermented bagasse.
Agave syrup as a substrate for inulinase production by Kluyveromyces marxianus NRRL Y-7571
Acta Scientiarum. Biological Sciences, 2016
The factorial planning was used to plan and optimize inulinase production by the yeast Kluyveromyces marxianus NRRL Y-7571. The experiments were conducted using a Central Composite Design (CCD) 22, at different concentrations of agave syrup (3.6 to 6.4%) and yeast extract (2.2 to 3.0%). After 96 hours of fermentation, the best condition for the inulinase production was 5% agave syrup and 2.5% yeast extract, which yielded an average of 129.21 U mL-1 of inulinase. Partial characterization of the crude enzyme showed that the optimal pH and temperature were 4.0 and 60°C, respectively. The enzyme showed thermal stability at 55°C for 4 hours.
Journal of Biotechnology, 2008
BACKGROUND: In Bacillus subtilis KCC103 α-amylase is hyper-produced and not catabolite repressed by glucose. Various sugars, raw starches and nitrogen sources were tested for their repression effect on α-amylase synthesis. Enhancement of α-amylase production by supplementing micronutrients and surfactants was studied. Using optimized medium, process parameters were optimized for improved α-amylase production.
Journal of Food Engineering, 2005
The inulinase production by yeast K. marxianus var. bulgaricus growing in yacon extract was investigated. The microorganism showed good development in yacon, higher enzymatic activities were achieved at 30% and 40% (v/v) of extract. The cultivation temperature (20, 25, 30, 35, 40°C) neither influenced the growth or the enzymatic activity. The optimum cultivation pH was 3.5. The highest activity was observed at 60°C and pH 4.0. At temperature of 55°C and 60°C occurred sharp decrease in the enzyme activity.
Electronic Journal of Biotechnology, 2013
Background: Inulinases have been extracted and characterized from inulin-storing tissues; however, production of microbial inulinases have recently draw much attention as they offer several industrial advantages. Many microorganisms, including filamentous fungi, yeast and bacteria have been claimed as inulinase producers. These hydrolases are usually inducible and their exo-acting forms may hydrolyze fructose polymers (inulin) and oligosaccharides such as sucrose and raffinose. Fungal inulinase extracts are often produced as stable mixture of highly active fructanhydrolases. From a practical prospective, the best known inulinases to date are those produced by species of Penicillium, Aspergillus and Kluyveromyces. Results: The production of extracellular inulinase by A. kawachii in liquid cultures, using either inulin or yacon derived materials as CES as well as inulinase inducers, is reported. In addition, a partial characterization of the enzyme activity is included. Conclusions: Yacon derived products, particularly yacon juice, added to the culture medium proved to be a good CES for fungal growth as well as an inducer of enzyme synthesis. Partial characterization of the enzyme revealed that it is quite stable in a wide range of pH and temperature. In addition, characterization of the reaction products revealed that this enzyme corresponds to an exo-type. These facts are promising considering its potential application in inulin hydrolysis for the production of high fructose syrups.
Production of inulinase from Kluyveromyces marxianus using Dahlia tuber extract
Brazilian Journal of Microbiology, 2012
Various carbon sources were evaluated for production of inulinase by yeast, Kluyveromyces marxianus MTCC 3995. Highest inulinase activity was observed with Dahlia extract (25.3 nkat mL-1) as carbon source. The enzyme activity was 1.4 folds higher than that observed in media containing pure chicory inulin (17.8 nkat mL-1). The yeast showed good growth on a simple medium containing dahlia extract (20% w/v) and yeast extract (2%w/v) as carbon and nitrogen source respectively, in 96 h. at 28°C and 120 rpm. Lowest inulinase yield (4.8 nkat mL-1) was seen in the medium containing glucose as C-source. Although varied inulinase levels were noticed on different C-sources, Inulinase: Sucrase (I/S) ratios were noticed to be similar. Among various protein sources tested, yeast extract was found to be the best source followed by beef extract (17.9 nkat mL-1) and peptone (13.8 nkat mL-1). The enzyme was optimally active at pH (4.0) and 50°C. TLC analysis of end product revealed that inulinase hydrolyzed inulin exclusively into fructose. Results suggest that the dahlia extract induced exoinulinase synthesis in Kluyveromyces marxianus and can be utilized as a potential substrate for inulinase production.
Food and Bioprocess …, 2009
In this work, production of inulinase was studied. Media formulation was optimized by experimental design and response surface techniques, as well as the pretreatment of the agro-industry residues used in the formulation of fermentation medium. Two agro-industry residues were investigated: sugarcane molasses (SCM) and corn steep liquor (CSL). Pretreatment with sulfuric acid was the most effective for clarification of SCM (pH 5.0, 24 h of resting time and final pH 4.0). Clarification of CSL was accomplished with phosphoric acid (pH 3.0, 24 h of resting time and final pH 5.5). A color reduction of approximately 70% was achieved for both substrates. The highest production of inulinase was obtained in a medium containing 100 g l −1 of pretreated SCM, 100 g l −1 of pretreated CSL and 6 g l −1 of Prodex Lac (yeast hydrolysate), yielding 1,139 U ml −1 .