PHYSICOCHEMICAL AND MICROBIOLOGICAL CHANGES ASSOCIATED WITH WINE PRODUCTION FROM PINEAPPLE USING PALM WINE YEAST

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ABSTRACTS

The physicochemical and microbiological changes associated with wine production with pineapple using palm wine yeast was studied. Pineapple juice was extracted and its sugar content was optimized before it was inoculated with yeast isolated from fresh palm wine and fermented for 11 days while the quality attributes and microbial biomass were determined every 48hours. The characteristics includes biomass build up, alcohol production, acidity levels, total solids, sugar content and specific gravity. Results obtained show that there was increase in the microbial biomass from 0.08g/L to 2.24g/L while alcohol production increased from 2.64 to 9.58%. The physiochemical properties also changed with fermentation time in which the pH decreased from 6.10 to 2.4 while titratable acidity increased from 0.33% to 2.39%. Sugar content decreased from 21.52% to 0.64% while total solid and specific gravity decreased from 18.82% to 6.37% and from 1.186 to 1.011 respectively. The temperature fluctuated between 30.00C and 32.00C. The fermentation dynamics was faster at the early days but slowed down after the 5th day. The observed changes in the physiochemical properties with fermentation time was attributed to the effect of the microbial activity during the fermentation while the increase in biomass was such as a proof that the yeast adapted well in the fermenting Must.




TABLE OF CONTENTS

Title Page                                                                                                                    i

Certification                                                                                                                ii

Dedication                                                                                                                   iii

Acknowledgement                                                                                                      iv

Table of contents                                                                                                        v

Lists of Tables                                                                                                            vii

Abstract                                                                                                                       viii

 

CHAPTER ONE

1.0        Introduction                                                                                              1

1.1       Aim                                                                                                                 8                     

1.2       Objectives                                                                                                       8

 

CHAPTER TWO

2.0       Literature Review                                                                                           9

2.1       Pineapple                                                                                                        12

 

CHAPTER THREE

3.0       Materials and Methods                                                                                   17

3.1       Collection of Samples                                                                                    17

3.2       Sample/Media Preparation                                                                             17

3.2.1    Preparation of Media for Yeast Isolation.                                                      17

3.2.2    Isolation of Palm Wine Yeast                                                                         18

3.3       Characterization of Yeast Isolate                                                                   18

3.4       Preparation of Pineapple Must                                                                       18

3.4.1    Determination of Sugar Content of Juice                                                       19

3.4.2    Optimization of Must                                                                                     20

3.5       Preparation of Inoculum                                                                                 20

3.6       Fermentation of Must                                                                                     20

3.7       Determination of Physicochemical Properties                                               21

3.7.1    Determination of temperature                                                                        21

3.7.2    Determination of pH                                                                                       21

3.7.3.   Determination of T. T. A                                                                                21

3.7.4    Determination of total solids                                                                          22

3.7.5    Determination of specific gravity                                                                   22

3.7.6    Determination of alcohol                                                                                23

3.7.7    Determination of biomass                                                                              23

3.7.8    Sensory Evaluation                                                                                         24

3.8       Statistical Analysis                                                                                         25

 

CHAPTER FOUR

4.0        Results                                                                                         26

 

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                              37

5.1       Discussion                                                                                                       37

5.2       Conclusion                                                                                                      39

5.3       Recommendations                                                                                          40

References                                                                                                      41

 

 

 

 

 

 

LIST OF TABLES

 

Table                                            Title                               Page

1                      Changes in quality characteristics of fermenting

Must during wine production                                                 27

 

2                      Quantitative Representation of the Panelist Score

Of Pineapple wine                                    28

 

 

 

 

 

LIST OF FIGURE


Figure                                     Title                                                       Page

 

1          Flow chart of pineapple wine production                                                                   16       

 

2          Changes in temperature of fermenting pineapple juice during wine production          29

 

3          Changes in pH of fermenting pineapple juice during wine production                   30

 

4          Changes in total solid in fermenting pineapple juice during wine production       31

 

5          Changes in TTA of fermenting pineapple juice during wine production                   32

 

6          Changes in sugar of fermenting pineapple juice during wine production                   33

 

7          Changes in specific gravity of fermenting pineapple juice during wine production          34

 

8          Changes in biomass in fermenting pineapple juice during wine production                   35

 

9          Changes in alcohol of fermenting pineapple juice during wine production                   36

 

 

 


 

 

 

                                                    CHAPTER ONE


1.0        INTRODUCTION

Fermentation was described by Prescott et al., (2008) as any process involving the mass culture of microorganisms either aerobically or anaerobically, and also the use of an organic substrate as the electron donor and the same partially degraded organic substrate as an electron acceptor. According to Pelezar et al., (2000), Louis Pasteur found out that fermentation of fruits and a grain resulting in alcohol production was brought about by microorganisms.

Fermentation is the metabolic conversion of a carbohydrate such as sugar into an alcohol or an acid using yeast, bacteria or a combination thereof (mixed culture) (Adams, 2005). It is also the slow decomposition of organic substance induced by microorganisms or by complex nitrogenous substances (Enzymes) of plant and animal origin (Board, 2003). In this process also, starch is broken down into fermentable sugars by fungal enzymes such as alpha amylase and glucoamylase to facilitate fermentation by yeast, mainly Saccharomyces species. Fermentation could occur under anaerobic or aerobic conditions and yields lactate, acetic acid, ethanol, carbon dioxide or some other simple product (Esechie, 2009). The numerous important products obtained by fermentation such as antibiotics, vitamins, feed supplements, and blood plasma expanders, has made fermentation as an industrial method for making specialty and various industrial chemicals gain wide attention (Horold et al., 2008). The production of industrial chemicals enhances the economic growth of any nation. Ethanol is one such industrial chemical used diversely as a solvent for chemicals, as an intermediate in the production of liquid detergents, in the manufacture of drugs, plastics, polishes, plasticizers and cosmetics, as antifreeze and as fuel ethanol (Akpan et al., 2005). Ethanol can be produced by Ethylene hydration or Fermentation (Agu et al., 2006).Ethanol can be made by fermentation using a number of renewable feed stocks, including sugar crops such as sugarcane, starch containing grains such as corn, or lignocelluloses materials including agricultural residues, herbaceous crops, and wood (Wang et al., 2004). Palm wine has been used locally in Nigeria for ethanol production by rural farmers. Palm wine is an alcoholic beverage obtained from the sap of various species of palm trees and is common in various parts of Africa. In Nigeria, it is abundant in the Niger delta and Rivers state in particular. The sap is extracted and collected by a tapper. Typically the sap is collected from the cut flower of the palm tree. A container is fastened to the flower stump to collect the sap. The white liquid that initially collects tends to be very sweet and non-alcoholic before it is fermented (Okafor, 2003).

Palm wine is a whitish, effervescent alcoholic beverage produced by the spontaneous fermentation of the sap of tree pals (family: Palmae). It is consumed in tropical and sub-tropical countries where palm grows.

These include South America, Asia and Africa (Blange and Bissir, 2009). It is refer to as toddy in Asian countries and tuba in South America. The type of palm which  the fermented sap is consumed varies in different parts of the world according to the type of palm wine obtained from oil palm (Elaeis guineensis), and raphia palm (Raphia vinifera) and Palmyra palm (Borassus flaellifer). Palm wine is a general name for alcoholic beverages produced from the sap of palm trees. It is different from the grape wines in that it is opaque. It is consumed all over the tropical world in Africa, Asia and South America. Palm wine is usually whitish and effervescent liquid both properties were derive from the fact that organisms are numerous and alive when the beverage is consumed (Okafor, 2004).

Palm wine differs from conventional beers and table wines produced in the modern brewery and winery in three ways. First, the media fermented for such beers are usually grains and for the wines, grapes and fruit juice are used in wine. The basic principle is however, is the same: a sugar solution is fermented, essentially by yeast (Saccharomyces species). Secondly, whereas there is control of fermentation during the production of modern  beers  and wine. Fermentation of palm wine is not controlled. Thirdly, the European beer and wines are usually clarified by removing microbial cells and other suspended material. Palm wine on the other hand is consumed without such clarifications. In essence, the basic differences between the true wines and the palm wine are a matter of technological differences between wine-making techniques and palm wine production methods. The methods of producing palm wine are likely to continue to change with enhanced technological advancement of the consuming countries (Benjakul,   et al., 2009).

During fermentation the yeast Saccharomyces cerevisiae produces a broad range of aroma-active substances, which are vital for the complex flavor of fermented beverages such as beer, wine and sake (Fleet et al., 2007). Flavour active substance produced by fermenting yeast cells can be divided into six groups, sulfur containing molecules, phenolic compounds and volatile esters (Saerens et al., 2010). Although volatile esters are only trace compounds in fermented beverages, they comprise the most important set of derived aroma active compounds. Volatile esters are of major industrial interest because they are responsible for the highly desired fruity, candy and perfume like aroma character of beer, wine, and sake (Fleet et al., 2007). Flavour in the most important distinguishing characteristics of most fermented foods, flavour is usually classified according to the source of different compound contributing to it. This include flavour contributed by the substrate prefermentative flavour (compound formed during extraction and conditioning of substrate), fermentative flavour (produced by yeast and bacteria during alcoholic and malolactic fermentation) and past, fermentative flavour (compound that appear during the storage period) (Orlica et al.,2007). Post fermentative flavour is usually a result of enzymatic. Is a complex phenomenon involving a number of factors in particular, it depends on the nature and concentration of the compounds initially present in the substrate, the capacity of the yeast to transform them and  the  fermentation  condition  employed.

Yeasts are eukaryotic microorganisms classified as fungi, with over 1,000 species currently known and described. It is believed that the species described so far represent only about 1% of all 1.5 million yeast species believed to exist on earth (Hutkins, 2006). Yeasts are unicellular fungus that reproduces either asexually by budding and transverse division (binary fission) or sexually through spore formation (Prescott, 2002). Although most yeast are unicellular, some species with yeast forms may become multi cellular through the formation of a string of connected budding cells known as pseudohyphae, or false hyphae, as seen in moulds. Yeast size can vary greatly depending on the species, typically measuring 3-4μm in diameter, although some yeast can reach over 40μm. The yeast species Saccharomyces cerevisae has been used in baking and fermenting alcoholic beverages for thousand years (Legrass et al., 2007).

Pineapple (Ananas satirus) is produced in large quantity in Nigeria. Madrid and Felice (2005) reported that about 4000 to 6000 tons of pineapple fruits are produced annually, most of them are wasted due to poor handling and poor storage methods. Fermenting pineapple juice goes a long way in reducing wastages and increasing local farmer’s income. The nutritional values of pineapple fruits according to Ihekoronye and Ngoddy (2007) are 54 mg/100 mL of vitamin C, 12% (w/v) of total sugars, 0.3% of protein, 0.1% of fats, 12 mg of calcium, 0.3 mg of iron, 0.08% of thiamine and 0.1 mg of niacin.

Yeast species are very useful in many industrial processes such as the production of alcoholic beverages, biomass and various other metabolic processes (Frazier and Westhoff, 2008). In baking and alcohol production, palm wine yeast (Saccharomyces cerevisiae) is most commonly used to convert sugars into ethanol and as the yeast utilizes the sugar, carbon dioxide and alcohol are produced (Zhou, 2002).

Wine production is a product of old biotechnology (Alian and Musenge, 2006), although the modern methods of production are in carefully controlling the processes in bioreactors. The juice from the crushed fruit is usually fermented by pure cultures of certain strains of yeast (Maldonado et al., 2005).

Processing steps includes crushing according to Querol et al. (2003).

The ability to produce palatable effervescent beverage by alcoholic fermentation of natural fruit juices is a demonstration of inherent ingenuity of man. The nutritional role of wine is important since its average contribution to total energy intake is estimated to be 10 to 20% in adult males (Macrae et al., 2011).

During the past few decades, grapes are the main fruits that were used for wine production. Despite that, several studies have investigated the suitability of other fruits as substrates for the purpose of wine production (Joshi and Bhutani, 2005; Joshi et al., 2003; Ndip et al., 2001; Okunowo et al., 2005).

Moreover, the non-availability of grapes, which is usually the fruit of choice for wine production in the tropics has necessitated the search for alternative fruit source in Nigeria and other tropical countries (Alobo and Offonry, 2009). In, Nigeria, there is abundance of tropical fruit which includes passion fruit, watermelon, pineapple, plum, orange etc., these fruits are highly perishable, and susceptible to bacterial and fungal contamination as a result they fail to reach the market due to spoilage, mechanical damage and over ripeness (Ihekoroye and Ngoddy, 2007). Besides, these fruits are difficult to keep for considerable length of time; hence the ripe fruits are utilized either as fresh or processed into juice and specialty products (Oyeleke and Olaniyan, 2007).

High rate wastage of these fruits especially at their peak of production during their season necessitates the need for alternative preservation food forms towards an enhanced utilization of these fruits. The production of wines from common fruits could help reduce the level of post harvest  losses and increase variety of wines (Okoro, 2007; Alobo and Offonry, 2009). Although, many genera and species of yeasts are found in musts, Saccharomyces cerevisae is the main yeast strain that is commonly reported to be responsible for alcoholic fermentation (Pretorius, 2000; Querol et al., 2003). However, many studies have investigated the use of other yeast strains (Ciani and Maccarelli, 2007; Okunowo et al., 2005) and also combination of yeast strains (Clemente-Jimenez et al., 2005) in fermentation especially in wine production. Reports have shown that the fermentation of fruit juices using yeast from different sources creates variety in flavour and varying levels of alcoholic contents in wines Clemente-Jimenez et al., (2005) in their work reported Pichia fermentans as good starter strains for most fermentation as Pichia fermentans in mixture with S. cerevisae improve the aroma as well as the characteristics features of the wine.

The fermentation of wine is known to be a complex process with various ecological and biochemical processes involving yeast strain (Fleet, 2003). Palm wine is a naturally sweet fermented beverage obtained from the sap of Elaeis species and the sap of Raphiaspp, which contains a heavy suspension of live yeasts and bacteria (Okafor, 2000). It is mostly common in Africa, especially Nigeria. Most studies on palm wine have reported its potentials as source of yeast isolate for the fermentation industries. Okafor (2004) in his study isolated seventeen yeast strains, four belonging to the species of Candida, twelve to the genus of Saccharomyces and one to Endomycopsis species. In grapes, yeast strains, such as Hanseniasporauvarum, Kloeckeraapiculata, Candida,Pichia, Rhodotorula and Kluyveromycesare known to be predominant (Duart et al., 2010).

The fermentation for the elaboration of beverage is known to depend on the performance of yeast to convert the sugars into alcohol and esters. Besides, the different species of yeast that develop during fermentation determine the characteristic of the flavour and aroma of the final product. Also, because different fruits have different composition, there is the need for yeast strains to adapt to different environments, such as sugar composition and concentration of acetic acid (Fleet, 2003; Duart et al., 2010). Although, tropical fruits and several yeast strains have been screened for their suitability in wine production, most studies have either focused only on the suitability of the fruits or the yeast strains.


1.1 AIM

The aim of this work was to produce wine with pineapple juice using yeast from palm wine.

 

1.2       OBJECTIVES

The objective includes specifically the following;

1.     To isolate and identify yeast from palm wine.

2.      To determine the changes associated with pineapple juice fermentation to produce wine.

 

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