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There are some differences in the anti-inflammatory activities of four typical components in EGB (extracts of ginkgo biloba leaves), and there is also a synergistic relationship. The order of inhibiting the NO-release ability of single functional components is OA > GF > OPC > G. Ginkgolide (G), proanthocyanidins (OPC), and organic acids (OA) all have synergistic effects on ginkgo flavonoids (GF). GF:OA (1:9) is the lowest interaction index among all complexes, showing the strongest synergy. The anti-inflammatory mechanism of the compound affects the expression of p-JNK, p-P38, and p-ERK1/2 proteins by inhibiting the expression of iNOS and COX2 genes on NFKB and MAPK pathways. This also provides a research basis for the development of anti-inflammatory deep-processing products of EGB.

Ginkgo biloba seed exocarp contains a variety of bioactive components, such as polysaccharides, flavonoids, terpenoid trilactones, and ginkgolic acids, which have a high value of utilization. However, Ginkgo biloba seed exocarp is always treated as waste of ginkgo seeds processing. To maximize the utilization of Ginkgo biloba seed exocarp, it is necessary to deliberate and arouse researchers' attention. This review demonstrated the extraction method, purification method, and determination method of bioactive components in Ginkgo biloba seed exocarp and summarized its composition and bioactivities. Moreover, the future study of Ginkgo biloba seed exocarp resource is prospected, hoping to provide a theoretical basis and direction for its further development and utilization., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

The predicted anti-oxidation is related to apoptosis, proliferation, lipid metabolism, cell differentiation, and immune response. There are some differences in the antioxidant capacity of the four typical components of ginkgo biloba extract (EGb) including ginkgo flavone (GF), ginkgolide (G), procyanidins (OPC), and organic acids (OA), and any two members of them can exhibit apparent synergistic effects. The order of DPPH scavenging ability was: OPC > GF > OA > G. The scavenging ability of procyanidins was close to that of VC; the scavenging capacity of ABTS was GF > OPC > OA > G. The GF:OPC (1:9) showed the best synergism in scavenging DPPH and ABTS radicals. The 193 kinds of small molecules reported in EGb were obtained by analyzing the properties of EGb. In order to construct a corresponding biological activity target set, molecular docking and the network pharmacology method were employed to build the molecular action mechanism network of a compound target, and the main biological functions and signaling pathways involved with their antioxidant activities were predicted. The results displayed that the top ten compounds which belonged to the two broad categories, ginkgo flavonoids and proanthocyanidins, could interact closely with several important target proteins (CASP3, SOD2, MAPK1, HSPA4, and NQO1). This would be expected to lay a theoretical foundation for the deep development of Ginkgo biloba extract.

4'- O -Methylpyridoxine (MPN) and MPN-5'-glucoside (MPNG) are collectively known as ginkgotoxin, which are the main toxic ingredients of excessive consumption of Ginkgo biloba seeds. Water extraction is the generally adopted sample preparation method for high-performance liquid chromatography determination of ginkgotoxin. However, endogenous enzymes such as glycosidases in Ginkgo biloba seeds can hydrolyze MPNG to MPN in the process of water extraction, which will result in the measured contents of MPN and MPNG but not their natural contents in Ginkgo biloba seeds. In this work, inhibitors for the endogenous enzymes were first screened, and it was found that silver fluoride could effectively inhibit endogenous enzymes such as glucosidase and phosphatase. The optimized concentration of silver fluoride was 25 mmol/L, which could effectively inhibit the endogenous enzymes for more than 60 h. A new sample preparation method based on water extraction with 25 mmol/L silver fluoride addition was thus developed. This method was employed to determine the native contents of MPN and MPNG in the exotesta and kernel of five Ginkgo biloba seed cultivars. The result showed that the contents of MPNG in the exotesta and kernel of five cultivars were significantly higher than those of MPN. MPNG was present at high content in raw seeds, which was the main form of ginkgotoxin in seeds. The method established in this work is simple and effective and can be used to accurately quantify the native contents of MPN and MPNG.

Ginkgo biloba L. is an ancient plant relic, which is known as a "living fossil", and is widely cultivated in China. This plant with medical potential and health benefits has drawn the attention of researchers. Ginkgo seeds are rich in protein. Ginkgo seed proteins (GSPs) have good functional properties over many other seed proteins, which have the potential to be utilized as food ingredients. Moreover, GSP contains no restricted amino acids and is easy to be separated. Several GSP isolate with various bioactivities, such as antimicrobial and antioxidative activities, have been purified and evaluated for their bioactive potential. In this review, the separation methods and bioactivities of GSP were summarized, physicochemical characteristics and functional properties were comprehensively reviewed and compared with other seed proteins. Some food applications of GSP were also briefly introduced. Besides, some suggestions and prospects were discussed in this review., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Ginkgo biloba seeds are well known for the significant curative effects on relieving cough and asthma. However, the development of products from ginkgo seeds still falls behind at present, resulting in a great waste of ginkgo seeds' resource. In this work, submerged fermentation of ginkgo seed powder using Eurotium cristatum was studied to investigate its feasibility as a new processing method., Results: To promote the growth of E. cristatum, the optimum fermentation medium was 80.0 g L -1 of ginkgo seed powder with addition of 5.0 g L -1 calcium chloride (CaCl 2 ), 4.0 g L -1 magnesium sulfate (MgSO 4 ), 1.25 g L -1 zinc sulfate (ZnSO 4 ) and 0.65 g L -1 iron(II) sulfate (FeSO 4 ). The optimum fermentation conditions were pH 5.8 ± 0.1, inoculum size 5.1 × 10 6  CFU mL -1 , liquid medium volume 100 mL in 250-mL Erlenmeyer flask and fermentation 4 days. Through fermentation, the production of lovastatin in fermentation broth could reach up to 32.97 ± 0.17 μg mL -1 and the total antioxidant capacity was improved by more than two-fold. In addition, 40.15% of the ginkgotoxin in ginkgo seed powder was degraded while the entire degradation of ginkgolic acids was obtained. Moreover, fermented ginkgo seed powder suspension presented pleasant fragrances, and the activities of amylase and protease were enhanced to 11.30 ± 0.10 U mL -1 and 23.01 ± 0.20 U mL -1 , respectively., Conclusions: Submerged fermentation using E. cristatum could significantly enhance the functional value and safety of ginkgo seed powder, and had great potential to become a novel processing method for the development of ginkgo seeds fermented products. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Background: Ginkgotoxin including 4'-O-methylpyridoxine (MPN) and MPN-5'-glucoside (MPNG) is responsible for Ginkgo seed food poisoning. The purpose of the work reported was to prepare detoxified Ginkgo seed powder and at the same time to retain the nutritional and functional components of Ginkgo seed powder to the maximum extent., Results: Resin adsorption technology was firstly employed to remove ginkgotoxin from water extract of Ginkgo seed powder. Under optimal conditions, the adsorption efficiency of the optimal resin for MPN could reach 100%, and that for MPNG could only reach 85.4 ± 0.93%. Resin adsorption alone could not effectively remove MPN and MPNG simultaneously. Endogenous enzymatic hydrolysis was next attempted to transform MPNG to MPN. MPNG could be completely hydrolyzed to MPN by endogenous enzyme(s) at 40 °C and pH 5.0 in 180 min. Ginkgotoxin only in the form of MPN in the enzymatic hydrolysate was then adsorbed with resin and the conditions were statistically optimized. The adsorption efficiency of MPN reached 98.89 ± 0.99% under the optimized conditions., Conclusions: Removal of ginkgotoxin by combining endogenous enzymatic hydrolysis with resin adsorption could preserve the main nutritional and functional components of Ginkgo seed powder to the most extent, and did not change its main characteristics. The ginkgotoxin removal method developed in this work is a relatively simple and efficient approach. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Ginkgo biloba leaves have various health benefits due to the presence of bioactive compounds such as polyprenyl acetates, flavonoids, and terpene trilactones. However, there is little literature reported on the aromatic acids in Ginkgo biloba leaves. In this work, five aromatic acids including shikimic acid (SA), 6-hydroxykynurenic acid (6-HKA), protocatechuic acid (PA), gallic acid (GAA), and p -hydroxybenzoic acid (PHBA) were simultaneously extracted from Ginkgo biloba leaves by employing the green deep eutectic solvents (DESs). A DES tailor-made from xylitol, glycolic acid and 1,5-pentanedioic acid at a molar ratio of 1:3:1 with 50% ( w / w ) water addition, named as NGG50, gave higher extraction yields for the five aromatic acids. Main factors affecting the extraction process were further optimized. The highest extraction yields of SA, GAA, 6-HKA, PA, and PHBA were 94.15 ± 0.96 mg/g, 332.69 ± 5.19 μg/g, 25.90 ± 0.61 μg/g, 429.89 ± 11.47 μg/g and 67.94 ± 0.37 μg/g, respectively. The NGG50-based extraction process developed here was a successful attempt of simultaneously extracting five aromatic acids from Ginkgo biloba leaves for the first time, which could provide a new exploitation direction of Ginkgo biloba leaves.

Proanthocyanidin (PAC) holds strong free radicals scavenging ability and is widely used as oxygen free radical scavenger. Huge demand, safety and economic aspect are forcing the PAC extraction to explore new resource and greener solvent. An efficient extraction method of PAC from Ginkgo biloba leaves was established by employing deep eutectic solvent (DES)-water mixture. After a rational design, a DES-water mixture (named as Ch-M 55 ) was chosen as the extraction solvent, which was prepared from choline chloride and malonic acid at a molar ratio of 1:2 with water addition of 55% (w/w). The main factors affecting the extraction yield were statistically optimized using a central composite design combined with response surface methodology (RSM). The optimal conditions were obtained as follows: extraction temperature of 65 °C, extraction time of 53 min and Ch-M 55 to solid ratio of 10.57:1 (v/w, mL/g). The PAC extraction yield was 22.19 ± 0.71 mg/g under the optimized conditions, which was much higher than those of conventional organic solvents. The antioxidant activity of PAC extracted by Ch-M 55 was similar to that of 70% acetone, but a little less than 70% methanol and 70% ethanol. The differences in constituent and polymerization degree of PAC extracted by different solvents might account for the difference in antioxidant activity. In consideration of biodegradability and pharmaceutical acceptability, the Ch-M 55 -based extraction method is obviously efficient, green and ecofriendly. Extraction of PAC from Ginkgo biloba leaves also provides a new PAC resource., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This paper presents a prefermentation treatment method involving fungi to improve flavonoid extraction from the leaves of Ginkgo biloba . The fungi employed for this treatment were screened from the soil present under an ancient ginkgo tree. Seventy-six strains belonging to 23 genera were isolated and identified by a molecular identification method employing 18S rDNA sequences. Thirty-three strains grew well using ginkgo leaves as the growth medium. One strain, Gyx086, with higher extracted yield of flavonoids and more similar to the control, was finally selected for prefermentation processing. The major fermentation factors were optimized by response surface methodology. The optimal conditions for the highest total falvonoid yield were 27.8 °C for temperature, 64.2% for moisture content, and 61 h for fermentation time. Under the optimal condition, a actual total flavonoid yield of 27.59 ± 0.52 mg/g dry weight culture sample was obtained, which was about 70% higher than that of unfermented gingko leaf samples.

Deep eutectic solvents (DESs)-based ultrasonic extraction of terpene trilactones (TTLs) fromGinkgo bilobaleaves was efficiently developed. Sixteen DESs were prepared, and DESs composed of choline chloride-urea (ChCl-U) and betaine-ethylene glycol (BE-EG) gave higher TTL extraction yields than the present, most efficient solvent 70% ethanol. The extraction conditions were further optimized, and the optimum conditions were as follows: taking BE-EG containing 40% (w/w) water as the extraction solvent, 1:10 ofG. bilobaleaves powder-to-solvent ratio, and ultrasonic treatment at 45°C and 100 W for 20 min. A total extraction yield of 1.94 ± 0.03 mg/g was obtained under the optimum conditions, which indicated that 99.37% of TTLs could be extracted from theG. bilobaleaves powder by a single extraction. Moreover, the polyamide resin was used to recover the TTLs in DES extracting solution, and recovery yield of 95.1% was attained. Therefore, BE-EG containing 40% (w/w) water was a potential alternative solvent for TTLs extraction fromG. bilobaleaves. [ABSTRACT FROM AUTHOR]