Microbiology and Chemical Characteristics of Tofu Waste-Based Probiotic Drinks

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‘‘Opportunities and Challenges for Sustainable Learning, Research and Community Service in Covid-19 Pandemic Constraints’

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Microbiology and Chemical Characteristics of Tofu Waste-Based Probiotic Drinks

Irmayanti^1*, Lukmanul Hakim², T. Makmur³, Paqrul Rizki¹,Chairuni AR¹

1Department of Agricultural Industrial Engineering, Faculty of Agricultural Technology, Serambi Mekkah University, Banda Aceh, Aceh, 23345, Indonesia

2Department of Food Technology, Faculty of Agricultural Technology, Serambi Mekkah University, Banda Aceh, Aceh, 23345, Indonesia

3Department of Agribusiness, Faculty of Agriculture, Syiah Kuala University, Banda Aceh, Aceh, 23111, Indonesia

*Corresponding Author: Irmayanti, [email protected]

Abstract

One industry that produces waste is tofu industry which produces solid waste and liquid waste from the washing, soaking, and cooking processes. To increase the added value of tofu industrial waste, tofu liquid waste can be used as a probiotic drink because it still contains relatively high protein in food processing.

Making probiotic drinks from tofu wastewater requires the addition of carbohydrates in the form of sucrose. This research used factorial completely randomized design(CRD) consisting of two factors. The first factor was the addition ofsucrose (S) consisting of 3 levels:S1 = 10%, S2 = 12%, S3 = 14%.The second factor was the concentration of lactobacillus casei consisting of 3 levels: L1 = 4%, L2 = 7%, L3 = 10%.The addition of sucrose (S) was found to have a very significant effect on the total microbes, total acid, but have no significant effect on thepH of tofu liquid waste probiotic drink. The best treatment of probiotic drinks was that resulting from the addition of 10% sucrose and a concentration of Lactobacillus casei 7% (S1L2) to produce good quality probiotic drinks with chemical properties, namely lactic acid bacteria 4.88, with pH of 3.57%, and total acid of 1.84%.

Keywords: probiotic, lactobacillus casei, sucrose, tofu waste.

1. Introduction

In the tofu making, solid and liquid waste are produced. However,both types of waste are often not properly managed and utilized. In fact, the waste still has high nutrition, such carbohydrates and protein. Solid waste can be reprocessed into various kinds of food, such as tempeh, flour, and cakes (Kaswinati, 2007), whereas liquid waste can be processed into biogas or re-sterilized before being discharged into the environment so that it will not harm the environment. In addition, tofu liquid waste can also be used optimally in probiotic drinks (Disyamto, 2014). Tofu liquid waste still contains high protein and is usable for the growth process of lactic acid bacteria in producing certain products, such as probiotic drinks (Fatma, 2012).

Probiotic drinks can be defined as drinks that are able to improve health because they contain live microorganisms, namely lactic acid bacteria (LAB) which can reach the digestive tract in an active condition (Wenny, 2015). Several types of lactic acid bacteria commonly

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81 used in the manufacture of probiotic drinksareBifidobacterium, Lactobacillus Bulgaricus, Lactobacillus Acidophiland Lactobacillus Casei because they have the ability to inhibit the growth of pathogenic bacteria(Dania et al., 2015).Lactobacillus are bacteria that can break down proteins, carbohydrates and fats in food, and help the absorption of important elements and nutrients, such as minerals, amino acids, and vitamins needed by humans and animals to survive (Fardiaz, 2014).

Making probiotic drinks from tofu wastewater requires the addition of carbohydrates in the form of sucrose. Apart from being a source of carbon and energy for LAB, sucrose is used in the manufacture of probiotic drinks as a source of energy for lactic acid bacteria and increases the antibacterial properties in probiotic drinks. The addition of sucrose will affect the growth of probiotic bacteria in beverages and their activity in utilizing carbohydrate components in the raw material composition for probiotic beverage products (Maryana, 2014). Kurniawati (2015) explains that 12% sucrose and 10% skim milk can produce pineapple skin juice probiotic drinks. In addition, Siadari (2014) states that the addition of 5%

sucrose and 15% skimmed milk provides good characteristics, and the amount of LAB is still quite high in sweet potato probiotic drinks.

2. Materials and Method

The basic ingredients in this study were tofu liquid waste from a tofu factory in Banda Aceh, Indonesia, lactic acid bacteria, namely Lactobacillus casei, sucrose, and aquades. This research was carried out using a factorial Completely Randomized Design (CRD) model consisting of 3 levels and 2 replications, namely the addition of sucrose (S) which consists of 3 levels, namely S1 = 10%, S2 = 12% and S3 = 14%, and the concentration of Lactobacilluscasei (L) which consists of 3 levels, namely L1 = 4%, L2 = 7% and L3 = 10%.

Production of Yogurt from Tofu Waste

The tofu liquid waste that has been filtered was put into an Erlenmeyer as much as 100 ml, then added with sucrose (10%, 12%, and 14%) and stirred until evenly distributed, then sterilized at 110 ° C for 10 minutes. The tofu liquid waste that has been sterilized was then cooled to a temperature of 45 ° C and the Lactobacillus casei starter was added (3%, 5%, and 7%) of the volume of liquid waste used, then fermented for 24 hours at 40 ° C to pH ± 4,5 fermentation was stopped and analyzed for total lactic acid bacteria, pH, acidity and organoleptic test.

Microbiology and Chemical Analysis of Yogurt from Tofu Waste

The parameters observed in this study include total acid (Umamet al., 2012), pH test (Wahyudi, 2006), and total lactic acid bacteria (Fardiaz, 1987).

3. Results and Discussion Lactic Acid Bacteria (LAB)

LAB is a bacterium capable of fermenting sugar or carbohydrates to produce lactic acid in large quantities. LAB analysis results from data can be seen in Table 1.

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82 Table 1. Data of Average Value of LAB Analysis Results for Probiotic Drinks Addition of sucrose (S) Concentration of Lactobacillus casei (L)

L1 = 4% L2 = 7% L3 = 10%

S1 = 10% 5.35 x 10⁶ 4.88 x 10⁶ 4.55 x 10⁶ S2 = 12% 5.38 x 10⁶ 5.35 x 10⁶ 5.36 x 10⁶ S3 = 14% 5.44 x 10⁶ 5.35 x 10⁶ 5, 45 x 10⁶

The highest LAB for probiotic drinks was obtained by adding 14% sucrose (S3) with the concentration of Lactobacillus casei 10 % (L3) was 5.45 x 10⁶. The lowest average LAB value was obtained by the addition of 14% sucrose (S3) and 10% Lactobacillus casei (L3) concentration of 4.55 x 10⁶.

Figure 1. Effect of the Addition of Sucrose to LAB of Probiotic Drinks (BNT0,01 = 0.63 and KK = 3.71).

From Figure 1, it can be seen that the higher concentration of sucrose added to the tofu liquid waste before fermentation increased the total LAB. This increase in total LAB occured because sucrose is a type of sugar that can be metabolized by LAB into lactic acid during the fermentation process. The addition of 10% sucrose and 4% Lactobacillus casei was the least addition of sucrose, but LAB was still growing.

The more sucrose available, the more substrate LAB can break down into pyruvic acid which can then be converted into other organic acids, such as lactic acid (Yunus & Zubaidah, 2015). Therefore, the more substrate available, the higher increase of LAB that grows. All

a [VALUE]

a 5.41

0 1 2 3 4 5 6

S1 = 10% S2 = 12% S3 = 14%

Total BAL Cfu/ml

Addition of sucrose (%)

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83 treatments that meet SNI 10-2981-2009 with an average amount of LAB 5.23 x 10⁶cfu / ml, because based on SNI 10-2981-2009, the amount of LAB produced in fermented drinks is a minimum of 1.0 x 10⁶ cells / ml. This is in accordance with the opinion of Winarno and Fernandez (2007) who state that the final fermentation product usually contains 1.0 x 10⁶cfu / ml.

pH (Degree of Acidity)

The highest pH of probiotic drinks was obtained at the addition of 14% sucrose (S3) and 4% (L1) Lactobacillus casei concentration of 4.06, while the lowest pH average value was obtained by the addition of 10% sucrose (S1) and 7% Lactobacillus casei concentration (L2) of 3.57.Data from the analysis of the pH of probiotic drinks can be seen in Table 2.

Table 2. Data of Average Value Analysis Results of pH analysis of probiotic drinks Addition of sucrose (S) Concentration Lactobacillus casei(L)

L1 = 4% L2 = 7% L3 = 10%

S1 = 10% 3.97 3.57 3.95

S2 = 12% 3.98 4.01 3.99

S3 = 14% 4.06 3.98 4.00

In terms of the addition of sucrose, the pH value for each treatment was not significantly different. Although not significantly different, the pH value of tofu liquid waste probiotic drink tends to decrease along with the increasing concentration of sucrose. When the LAB fermentation process occurred, LAB produced lactic acid so that the more lactic acid produced tends to decreasethe pH.

Total Lactic Acid

Total lactic acid is a measurement of all acid components, including dissociated ones.

Besides lactic acid, there are also other easily dissociated acids, such as acetic acid and formic acid (Hernawati, 2002).Data from the analysis of the total acid probiotic drink can be seen in Table 3.

Table 3. Data of Average Value Analysis Results of the Total Acid Probiotic Drink Addition of sucrose (S) Concentration Lactobacillus casei(L)

L1 = 4% L2 = 7% L3 = 10%

S1 = 10% 1.77 1.84 1.77

S2 = 12% 1.60 1.69 1.64

S3 = 14% 1.50 1.67 1.70

The highest total acid probiotic drink was obtained at the addition of 10% sucrose (S1) and 7% (L2) Lactobacillus casei concentration of 1.84%, while the lowest average total acid

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84 value was obtained by the addition of 14% sucrose (S3) and the concentration of 4 % (L1) of 1.50% of Lactobacillus casei.

Figure 2. The Effect of Adding Sucrose to the Total Lactic Acid of Probiotic Drinks (BNT0.01 = 0.20 and KK = 3.74).

From Figure 2, it can be seen that the higher the addition of sucrose, the more total acid value in probiotic drinks decreased. The value of the lactic acid content was inversely proportional to the pH value. The production of lactic acid by lactic acid bacteria causes a decrease in the acidity of probiotic drinks during the addition of l sucrose. The decrease in the acidity of probiotic drinks may be due to the growth of microorganisms that were not too good. This is in accordance with the opinion of Hadiwiyoto (2008) who states that the high and low levels of lactic acid in fermented milk products are influenced by the number and type of starter used. The buffering effect may be one of the factors for this to occur.

Additionally, the influence of other alkaline substances that were able to neutralize the acids contained in probiotic drinks such as protein, carbohydrates, and dissolved solids also caused the lactic acid content in probiotic drinks low. Furthermore, probiotic drinks have limited levels of protein and nutrients, so it is necessary to add peptone for optimal microorganism growth.

a [VALUE]

b [VALUE]

a [VALUE]

0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 2

S1 = 10% S2 = 12% S3 = 14%

Total Lactic Acid(%)

Addition of sucrose (%)

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85 Figure 3. Effect of the Concentration of Lactobacillus Casei on the Total Lactic Acid of

Probiotic Drinks (BNT0,01 = 0.12 and KK = 3,74).

From Figure 3, it can be seen that the higher the concentration of Lactobacillus casei, the more total lactic acid produced tends to decrease. In the treatment, the concentration of addition of lactic acid bacteria was 4%, and the amount of lactic acid was not too high;however, the increase in lactic acid was quite high in the addition of 7% concentration, but slightly decreased at the concentration of addition of 10%. This was presumably because the addition of bacteria was still relatively small. The higher the concentration of bacteria added, the higher the number of lactic acid bacteria which can hydrolysed carbohydrates to lactic acid and which can increase the ability of bacteria to break down lactose in skim milk added to tofu liquid waste. This shows that the pH of probiotic drinks decreasedwith the increasing addition concentration of lactic acid bacteria.

4. Conclusion

The addition of sucrose (S) had a very significant effect on total microbes and total acid but had no significant effect on pH of tofu liquid waste probiotic drink. The best treatment of probiotic drinks was that resulting from the addition of 10% sucrose and a concentration of Lactobacillus casei 7% (S1L2) to produce good quality probiotic drinks with chemical properties, namely lactic acid bacteria 4.88, pH 3.57%, and total acid 1.84%.

a [VALUE]

b [VALUE]

0 0,2 0,4 0,6 0,8 1 1,2 1,4 1,6 1,8 2

L1 = 4% L2 = 7% L3 = 10%

Total Acid (%)

ConsentrationLactobacillus casei(%)

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