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Bitterness in citrus juice is a common quality issue in the fruit-based beverage industry. Naringin, the dominant flavonoid responsible for bitterness in citrus fruit juice, can be mitigated by the enzyme naringinase. This study focuses on the isolation of novel naringinase-producing bacterium from mandarin orange peel, followed by the production and partial purification of naringinase. The bacterial strain exhibiting the highest enzyme production potential was identified as Bacillus megaterium AULS 1. The effects of carbon source (naringin) concentration, inducer (naringenin), pH, and temperature on naringinase production were investigated. Optimized conditions yielded a maximum naringinase production of 386.43 U mL–1. The produced enzyme was partially purified using ammonium sulfate precipitation and dialysis. The effects of pH, temperature, and metal ion concentration on the activity of the partially purified enzyme were evaluated, resulting in a highest activity of naringinase of 539.25 U mL–1 under optimized conditions. The debittering activity of the partially purified enzyme was 45.78 %. These findings demonstrate that Bacillus megaterium AULS 1 has the potential for naringinase production, and can be utilized for debittering citrus juices.

Yuzu (Citrus junos Sieb.) is a peel-edible fruit with a pleasant aroma, but its bitter taste can impact consumer appeal. In this study, an efficient enzymatic method reduced bitterness in green yuzu powder (GYP). Cellulase KN and naringinase from Aspergillus oryzae NYO-2 significantly decreased naringin and neohesperidin content by over 87 %, while increasing total dietary fiber and soluble dietary fiber by up to 10 % and 51 %, respectively. Insoluble dietary fiber decreased by up to 22 %. Cellulose, hemicellulose, lignin, and pectin contents in enzyme-treated YP decreased by 1.15–2.00-fold, respectively. Enzyme-treated GYP exhibited improved physicochemical properties, including enhanced solubility, oil-holding capacity, and water swelling capacities. 3T3-L1 cells treated with cellulase-treated GYP and naringinase-treated GYP showed lower lipid accumulation and higher lipolysis capability than GYP, along with decreased fatty acid synthase contents. These findings suggest that enzyme-treated GYP holds potential as a functional ingredient in the food industry.

The application of Neurospora sp., a fungus that commonly thrives on complex agricultural and plant wastes, has proven successful in utilizing citrus peel waste as a source of naringin. A UV-Vis spectrophotometric method proved the biotransformation of naringin, with an absorption maximum (λmax) observed at 310 nm for the biotransformed product, naringenin (NAR). Further verification of the conversion of naringin was provided through thin layer chromatography (TLC). The Neurospora crassa mediated biotransformation of naringin to NAR was utilized for the rapid (within 5 min) synthesis of silver (Ag) and gold (Au) nanoconjugates using sunlight to accelerate the reaction. The synthesized NAR-nano Ag and NAR-nano Au conjugates exhibited monodispersed spherical and spherical as well as polygonal shaped particles, respectively. Both of the nanoconjugates showed average particle sizes of less than 90 nm from TEM analysis. The NAR-Ag and NAR-Au nanoconjugates displayed potential enhancement of the antimicrobial activities, including antibacterial and nematicidal properties over either standalone NAR or Ag or Au NPs. This study reveals the potential of naringinase-producing Neurospora sp. for transforming naringin into NAR. Additionally, the resulting NAR-Ag and NAR-Au nanoconjugates showed promise as sustainable antibiotics and biochemical nematicides.

This study aimed to develop a method of naringinase biosynthesis by Aspergillus niger KMS on an optimized culture medium. The concentration of the six medium components in shake flasks was optimized by the Box and Wilson factor gradient method. Naringinase’s substrate, naringin, powdered albedo, flavedo, and red grapefruit segment membranes were used to stimulate naringinase biosynthesis. Rhamnose was chosen as the carbon source, while the nitrogen source was yeast extract and sodium nitrate. Naringinase biosynthesis was most favorable in the culture medium with the following composition (g 100 mL): 3.332—NaNO3; 3.427—yeast extract; 0.184—KH2PO4; 0.855—red grapefruit albedo; 0.168—naringin; 2.789—rhamnose. The obtained Aspergillus niger KMS culture fluid was concentrated, thereby precipitating the protein. As a result, a naringinase preparation with high activity, equal to 816 µmol × min−1 × g−1, was obtained.

Naringin and limonin are the two main bitter compounds of citrus products such as grapefruit juice. The aim of this investigation was to evaluate the reduction in both bitter components simultaneously using a combined biochemical and physical approach. The proposed strategy was based on the use of heterofunctional supports with glyoxyl groups that allow for the covalent immobilization of naringinase, which hydrolyses naringin and alkyl groups that allow for the adsorption of limonin. The supports were butyl-glyoxyl agarose (BGA) and octyl-glyoxyl agarose (OGA), which were characterized in terms of aldehyde group quantification and FTIR analysis. The optimal pH and temperature of free and immobilized enzymes were assessed. The maximum enzyme loading capacity of supports was analyzed. Debittering of grapefruit juice was evaluated using soluble enzyme, enzyme-free supports, and immobilized catalysts. Enzyme immobilized in BGA reduced naringin and limonin concentrations by 54 and 100%, respectively, while the use of catalyst immobilized in OGA allowed a reduction of 74 and 76%, respectively, obtaining a final concentration of both bitter components under their detection threshold. The use of OGA biocatalyst presented better results than when soluble enzyme or enzyme-free support was utilized. Biocatalyst was successfully applied in juice debittering in five repeated batches.

Naringinase is an enzymatic complex used in the deglycosylation of compounds with a high application potential in the food and pharmaceutical industries. The aim of the study was to immobilize naringinase from Aspergillus niger KMS on a magnetic carrier obtained on the basis of carob gum activated by polyethyleneimine. Response surface methodology was used to optimize naringinase immobilization taking into account the following factors: pH, immobilization time, initial concentration of naringinase and immobilization temperature. The adsorption of the enzyme on a magnetic carrier was a reversible process. The binding force of naringinase was increased by crosslinking the enzyme with the carrier using dextran aldehyde. The crosslinked enzyme had better stability in an acidic environment and at a higher temperature compared to the free form. The immobilization and stabilization of naringinase by dextran aldehyde on the magnetic polysaccharide carrier lowered the activation energy, thus increasing the catalytic capacity of the investigated enzyme and increasing the activation energy of the thermal deactivation process, which confirms higher stability of the immobilized enzyme in comparison with free naringinase. The preparation of crosslinked naringinase retained over 80% of its initial activity after 10 runs of naringin hydrolysis from fresh and model grapefruit juice.

Naringinase is an enzyme complex which exhibits α-l-rhamnosidase and β-d-glucosidase activity. This enzymatic complex catalyzes the hydrolysis of naringin (4′,5,7-trihydroxy flavanone 7-rhamnoglucoside), the main bittering component in grapefruit. Reduction of the level of this substance during the processing of juice has been the focus of many studies. The aim of the study was the immobilization of naringinase on chitosan microspheres activated with glutaraldehyde and, finally, the use of such immobilized enzyme for debittering grapefruit juice. The effect of naringinase concentration and characterization of the immobilized enzyme compared to the soluble enzyme were investigated. The maximum activity was observed at optimum pH 4.0 for both free and immobilized naringinase. However, the optimum temperature was shifted from 70 to 40 °C upon immobilization. The KM value of the immobilized naringinase was higher than that of soluble naringinase. The immobilization did not change the thermal stability of the enzyme. The immobilized naringinase had good operational stability. This preparation retained 88.1 ± 2.8% of its initial activity after ten runs of naringin hydrolysis from fresh grapefruit juice. The results indicate that naringinase immobilized on chitosan has potential applicability for debittering and improving the sensory properties of grapefruit juices.

Brazil is the world's largest producer of orange and concentrated juice for export. Concentrated juice with high levels of naringin has excessive bitterness, which reduces the quality and value on the market. The debittering can be obtained by using naringinase, an enzymatic complex that degrades naringin. This study reports the production of naringinase by Aspergillus niger 426 utilizing both sugar cane molasses as carbon source and yeast extract as nitrogen source. Naringin was used as inducer. Five nitrogen sources were studied and yeast extract was found as the best one as 225.6 mU/mL of naringinase activity at 120 hours of fermentation was achivied. For optimization of naringinase production it was applied response-surface methodology, with 22 incomplete factorial design. An activity value of 354.26 mU/mL of naringinase was achivied when as independent variables yeast extract (14.0g/L) and naringin (0.2g/L) were used. When applied ultrasound waves at 20 kHz of intensity for 2 minutes in the fermented broth, the activity reached the highest value of 473.6 mU/mL. Thus, naringinase, this enzyme of great potential for biotechnological applications, has its production increased by using statistical design and ultrasound.O Brasil é o maior produtor mundial de laranja e de suco concentrado destinado à exportação. Sucos cítricos concentrados para exportação com níveis elevados de naringina são excessivamente amargos, o que reduz a qualidade e o valor comercial do produto. A redução do amargor pode ser obtida pela naringinase, um complexo enzimático que degrada a naringina. Neste trabalho Aspergillus niger 426 foi usado como produtor de naringinase utilizando matérias-prima da agroindústria, melaço como fonte de carbono e extrato de levedura como fonte de nitrogênio. Naringina foi usada como indutor. Dentre as cinco fontes de nitrogênio pesquisadas, extrato de levedura apresentou o melhor resultado de 225,6 mU/mL de naringinase em 120 horas de cultivo. Para otimização da produção de naringinase, foi aplicada a metodologia de superfície de resposta, com planejamento fatorial incompleto 22, obtendo 354,26 mU/mL de atividade de naringinase, tendo como variáveis independentes o extrato de levedura (14,0g/L) e naringina (0,2g/L). Quando aplicadas ondas de ultrassom com intensidade de 20 kHz por 2 minutos na cultura, a atividade de naringinase atingiu o valor máximo de 473,6 mU/mL. Assim, a naringinase, enzima de grande potencial de aplicação biotecnológica, teve a produção aumentada pelo uso do planejamento estatístico e ultrassom.

The goal of this research is debittering of orange juices with enzimatic process. Naringinase enzim is produced with Penicillium decumbens. The research metode is complete random design with two variabel. The first variabel is concentration of enzim : 0,0; 200; 400 ppm and the second variabel is time of incubation : 1, 2 and 3 hours. The product of this research is tested ascorbic acid, Total Plate Count and organoleptic test. The result showed concentration of naringinase enzim 400 ppm and incubation 3 hours can eliminated the after taste of orangge juice without reduce the ascorbic acid and colour of orange juice.