Original Article

Assessment of the Process Variables for Degradation of Oxalate by Lactobacillus acidophilus ATCC 4356 Using Simulated Rumen Fluid Media and Tea

Dina Karamad, Kianoush Khosravi-Darani, Hedayat Hosseini, Sanaz Tavasoli, Aaron W. Miller

Applied Food Biotechnology, Vol. 7 No. 4 (2020), 18 August 2020, Page 195-204


Background and Objective: Lactobacillus acidophilus ATCC 4356 was used for oxalate degradation activity. In the present study, for the first time, the simultaneous influence of process variables on the analysis of high oxalate concentration and its optimization in the simulated intestinal environment was performed. In the end, the optimal results were performed in a tea environment as a common oxalate-containing beverage.

Materials and Methods: After screening the design of ten process variables including pH, glucose, sucrose, inulin, ammonium, sodium oxalate, yeast extract, sodium acetate, inoculum age, and size using Plackett-Burman design, a Box-Behnken method was used with four major variables of pH, glucose, sodium oxalate, and inulin

Results and Conclusion: Results showed that oxalate degradation in simulated rumen fluid was significantly affected by pH and sodium oxalate and glucose concentrations. At optimized conditions, oxalate degradation reached 48.94 ±0.98% of initial concentration. Furthermore, oxalate degradation was investigated in tea (as the most common hot drink in many countries such as Iran) at various times, temperature, and glucose concentration. At optimum condition, oxalate concentration reached 98.86% ±1.05 (from 264 to 24 mg per100 ml).

Improvement of Astaxanthin Production in Coelastrum sp. by Optimization Using Taguchi Method

Ameerah Tharek, Adibah Yahya, Madihah Md Salleh, Haryati Jamaluddin, Shinji Yoshizaki, Rozzeta Dolah, Hirofumi Hara, Koji Iwamoto, Shaza Eva Mohamad

Applied Food Biotechnology, Vol. 7 No. 4 (2020), 18 August 2020, Page 205-214


Background and Objective: Astaxanthin is a keto-carotenoid pigment known as one of the most valuable compounds with great potentials in the market. It has widely been used in nutraceutical, pharmaceutical, cosmetics and food industries due to its strong antioxidant activity. Green microalgae seem as promising natural sources in production of astaxanthin. The aim of this study was to optimize astaxanthin production in Coelastrum sp. to overcome low productivity of microalgae.

Materials and Methods: This study was carried out using experimentally statistical technique and Taguchi method to find optimum conditions for maximizing production of astaxanthin in green microalgae, Coelastrum sp. Effects of nutritional (carbon and nitrogen) and environmental (light and salinity) factors on biomass and astaxanthin production were investigated. Experiments were carried out for light intensity (250-550 µmol photons m-2 s-1), salinity using sodium chloride (1.0-3.0 g l-1), carbon source using sodium acetate
(0.5-2.0 g l-1) and nitrogen source using sodium nitrate (0.1-0.3 g l-1).

Results and Conclusion: Results showed that optimum conditions of astaxanthin production in Coelastrum sp. included 250 µmol photons m-2 s-1 of light intensity, 3 g l-1 salinity, 0.5 g l-1 carbon and 0.1 g l-1 nitrogen with a maximum yield of astaxanthin (14.44 mg l-1), which was 2-fold higher than that before optimization. This optimization resulted in high quantities of astaxanthin production using optimization of conditions that affected production yields of astaxanthin from Coelastrum sp.

Immobilization Effects of Wheat Bran on Enhanced Viability of Dairy Starters under Acid and Bile Salts Stresses

Yelena Oleinikova, Alma Amangeldi, Makpal Yelubaeva, Aigul Alybaeva, Sadanov Amankeldy, Margarita Saubenova, Anna Chizhaeva, Aida Aitzhanova, Ramza Berzhanova

Applied Food Biotechnology, Vol. 7 No. 4 (2020), 18 August 2020, Page 215-223


Background and Objective: Survival of beneficial microorganisms in human gut faces many challenges. Immobilization on dietary fibers not only increases the viability of probiotic cultures, but also improves intestinal microbiota composition and decreases several diseases. Therefore, the objective of this study was to assess effects of wheat bran immobilization on survival of multiple species dairy starters under high acidity and bile salts conditions.

Materials and Methods: Dairy starter association included lactic acid bacteria of Lactobacillus delbrueckii RKM 0850, Lactobacillus paracasei RKM 0852 and Lactobacillus. parabuchneri RKM 0854, acetic acid bacteria of Acetobacter syzygii RKM 0855, propionic acid bacteria of Propionibacterium freudenreichii subsp. shermanii PL and yeast of Kluyveromyces marxianus RKM 0853. Physical immobilization of the whole association was carried out using adsorption on 3% wheat bran.

Results and Conclusion: Effects of acid and bile salts stresses on microbial association survival largely depended on the taxonomic affiliation, aeration rate and experiment media. Kluyveromyces marxianus RKM 0853 was highly resistant to pH 2.0-3.0 and addition of 0.5-15.0% of bile salts. Furthermore, Lactobacillus paracasei RKM 0852 and Acetobacter syzygii RKM 0855 were the most resistant microorganisms to bile salts and Lactobacillus delbrueckii RKM 0850 and Lactobacillus parabuchneri RKM 0854 included the best survival rates at low pH. Wheat bran immobilization promoted survival of the microbial association under stress conditions and prevented loss of viability by more sensitive species. These results are essential for understanding and improvement of stress tolerance in probiotic microorganisms of various taxonomies.

Photoautotrophic Cultivation of Arthrospira maxima for Protein Accumulation under Minimum Nutrient Availability

Wanida Pan-utai, Natapas Poopat, Pramuk Parakulsuksatid

Applied Food Biotechnology, Vol. 7 No. 4 (2020), 18 August 2020, Page 225-234


Background and Objective: Arthrospira cyanobacteria are important freshwater blue-green microalgae as food protein sources for humans. Arthrospira maxima phototrophic cultivation was carried out under various minimum media formulations to enhance biomass and protein productions. Macronutrients and micronutrients are vital for biomass production and protein accumulation of Arthrospira maxima for food supplemented uses.

Materials and Methods: Photoautotrophic cultivation of Arthrospira maxima IFRPD 1183 was carried out using various Zarrouk culture media and inoculum preparation conditions for biomass and protein productions. Arthrospira maxima IFRPD 1183 was cultivated using algal chamber under closed and open photobioreactor systems.

Results and Conclusion: Micronutrients of B6 solution; NH4VO3, K3Cr2 (SO4)4. 24H2O, NiSO4. 7H2O, Na2WO3, Co (NO3)2. 6H2O and Ti2 (SO4)3 did not affect cell growth and protein accumul-ation in absence of media for inoculum production. Inoculum preparation under various conditions of Arthrospira maxima IFRPD 1183 was studied using filtration of the old media before use (cell filtration inoculum) with no filtrations of the old media before use (cell non-filtration inoculum). Cell non-filtration inoculum preparation was reported appropriate for biomass production and protein accumulation. For large-scale productions, open pond system of Arthrospira maxima IFRPD 1183 resulted in maximum biomass and protein production at nearly 1 g l-1 and 64% (DW), respectively, for repeat batch photoautotrophic cultivation. Absence of micronutrients with cell non-filtration inoculum was reported as an easy process to achieve large biomass and protein productions for closed and open photoautotrophic cultivations of Arthrospira maxima at decreased costs.


Background and Objective: Enzyme-assisted hydrolysis of the proteins from food sources is an effective way to generate peptides with various bioactive properties. Furthermore, enzyme immobilization is a way to recycle enzymes for the future uses. The objective of this study was to investigate the effects of neutrase and thermolysin immobilization on the enzyme properties and in vitro anti-diabetic properties of intestinal digests achieved by the simulated digestion of egg white protein hydrolysates.

Material and Methods: Neutrase and thermolysin were immobilized on cellulose-coated magnetite nanoparticles. Then, enzyme activity, thermal resistance, reusability and optimum conditions of the egg white protein hydrolysis were assessed. Egg white protein hydrolysates were then digested in vitro and inhibitory activities of the intestinal digests against dipeptidyl peptidase IV (DPP-IV) and α-glucosidase were investigated.

Results and Conclusion: Enzymes immobilization resulted in increases in the thermal stabilities of them. Optimum temperatures for the egg white protein hydrolysis increased by 4.0 and 3.2 °C for neutrase and thermolysin, respectively. Digests from the hydrolysates of free neutrase effectively inhibited DPP-IV and α-glucosidase by 17.9 and 29.7%, respectively. These values for the hydrolysates released by the free thermolysin were higher (37.2 and 35.1%, respectively). The enzyme immobilization resulted in a 4.4% decrease in DPP-IV inhibitory activities of the digests for the hydrolysates from neutrase and a 28.6% decrease for those from thermolysin. Decreases in α-glucosidase inhibition due to the immobilization included 9.8% for neutrase and 12.2% for thermolysin for the digests from the hydrolysates. Based on the results from the current study, hydrolysates from the egg white proteins achieved by the free and immobilized neutrase and thermolysin can be used in formulations of the functional foods and nutraceuticals with multifunctional properties.

Optimization of Soy-based Media for the Production of Biologically Active Exopolysaccharides by Medicinal Mushroom Trametes versicolor

Hale Alvandi, Maryam Ghahremani, Ashrafalsadat Hatamian-Zarmi, Bahman Ebrahimi Hosseinzadeh, Zahra Beagom Mokhtari-Hosseini, Seyedeh Nazila Jafari Farjam

Applied Food Biotechnology, Vol. 7 No. 4 (2020), 18 August 2020, Page 251-261


Background and Objective: Diabetes mellitus is the fifth leading cause of death in the world. Damaging effects of diabetes include advanced metabolic complications and various organ lesions. More than 90% of diabetic cases belong to type 2 diabetes. For decades, medicinal fungi such as Trametes versicolor have been considered as treatments for diabetes. The fungi exopolysaccharides show α-glucosidase activity in the intestinal membrane.

Material and Methods: In this study, Iranian Trametes versicolor was cultured in various media based on soy milk, cow milk and soy protein extraction. After selecting appropriate culture media for biomass and exopolysaccharide production, response surface method with Box-Behnken design was used to investigate effects of independent variables of glucose (g l-1), soy milk (v v-1) and pH on biomass and exopolysaccharide production and optimization of these products. An animal (rat) model of streptozotocin-induced diabetic rats was used to investigate effects of exopolysaccharides on diabetes. Rats were treated with exopolysaccharides for 21 days.

Results and Conclusion: Results showed that the culture media containing soy milk was appropriate for the growth and production of fungal products. Productions of biomass and exopolysaccharides increased to 2.87 g l-1 and 1.37 g l-1 respectively. Based on the analysis of response surface method, pH with p-value of 0.0004 and pH and soybean interaction with p-value of 0.0129 included significant effects on exopolysaccharide production. In optimum condition of the culture media with pH 4.67, 13.01 g l-1 glucose and 75% v v-1 soybean milk, biomass and exopolysaccharides reached 21.80 g l-1 and 9.6 g l-1. In diabetic rat model treated with exopolysaccharides, a 50.38% decrease in blood glucose was seen while triglycerides, cholesterol and low-density lipoproteins included 89, 20 and 21.67%, respectively. Furthermore, high-density lipoproteins increased by nearly 2.5%.

Pullulan Production Using Molasses and Corn Steep Liquor as Agroindustrial Wastes: Physiochemical, Thermal and Rheological Properties

Saman Zarei, Faramarz Khodaiyan, Seyed Saeid Hosseini, John F. Kennedy

Applied Food Biotechnology, Vol. 7 No. 4 (2020), 18 August 2020, Page 263-272


Background and Objective: Pullulan is a microbial exopolysaccharide with wide uses in various industries. The aim of this study was to investigate pullulan production from agro-industrial wastes and study of pH, molasses concentration and corn steep liquor concentration as independent variables and yield of pullulan as response.

Material and Methods: Briefly, 5% (v v-1) of the inoculation media (yeast extract 3 g, malt extract 3 g, peptone 5 g and sucrose 10 g per liter of distilled water), including Aureobasidium pullulans were added into media, containing 100 ml of molasses (100, 150 and 200 g l-1) and various corn steep liquor concentrations (20, 40 and 60 ml l-1) at adjusted pH (4.5, 5.5 and 6.5). After extraction and separation of the biomass using centrifuge, two folds of the supernatant volume of cold ethanol were added to the samples and stored at 4 °C for 24 h. After centrifuging, pullulan was dried and analyzed using Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis and rheological tests.

Results and Conclusion: Findings revealed that the maximum production yield (18.29 g l- 1 ±0.10) was achieved under optimum fermentation conditions (pH of 5.3, molasses concen-tration of 165 g l-1 and corn steep liquor concentration of 43 ml l-1). Then, physiochemical and thermal properties of the pullulan under the highlighted conditions were investigated. Pullulan included 78.8% solubility with no hygroscopicity. Furthermore, structural analysis using Fou-rier transform infrared and X-ray diffraction verified presence of pullulan with an amorphous structure in the supernatant. The exopolysaccharide included acceptable thermal stability and gel-like behavior; in which, the elastic component was predominant based on the results of thermogravimetric analysis and rheological properties, respectively.


Background and Objective: Fish tissue and liver and some microalgae are some of the richest sources of omega-3 (ω3) fatty acids in nature. Annually, more than 4.2 billion tons of algae are cultivated with various goals. Therefore, development of novel ways to extract more ω3-fatty acids from microalgal tissues is a valuable target. In this study, comparisons were carried out between various methods of extraction and purification of ω3-fatty acids from a native species of microalgae.

Material and Methods: Combination method (urea complex and winterization) used as a method for extraction of ω3-fatty acids. For the optimization of combination method, three factors, including temperature of winterization, time of winterization and frequency of extraction with hexane were studied at four various levels and 15 examinations using Design Expert 10.0 software.

Results and Conclusion: Results of optimization included 10 h for time of winterization,
-20 °C for temperature of winterization and six times of extraction with hexane. In optimized conditions, the highest quantity of ω3-fatty acids derived from Chlorella vulgaris at the moisture of 65% was 43.6 mg g-1 fatty acids.