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Z.Z. Ni College of Food and Bioengineering, Xuzhou University of Technology, 221018, Xuzhou, Jiangsu Province, China

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J.T. Li College of Food and Bioengineering, Xuzhou University of Technology, 221018, Xuzhou, Jiangsu Province, China

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S.M. Zhang College of Food and Bioengineering, Xuzhou University of Technology, 221018, Xuzhou, Jiangsu Province, China

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Y.W. Dong College of Food and Bioengineering, Xuzhou University of Technology, 221018, Xuzhou, Jiangsu Province, China

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Q.Q. Liu Key Laboratory for Bioactive Materials of the Ministry of Education, Institute of Molecular Biology, College of Life Science, Nankai University, 300071 Tianjin, China

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M.G. Li Key Laboratory for Bioactive Materials of the Ministry of Education, Institute of Molecular Biology, College of Life Science, Nankai University, 300071 Tianjin, China

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Y.N. Wang College of Food and Bioengineering, Xuzhou University of Technology, 221018, Xuzhou, Jiangsu Province, China

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Abstract

Nattokinase (NK) is effective in the prevention and treatment of cardiovascular disease. Cucumber is rich in nutrients with low sugar content and is safe for consumption. The aim of this study was to construct a therapeutic cucumber that can express NK, which can prevent and alleviate cardiovascular diseases by consumption. Because the Bitter fruit (Bt) gene contributes to bitter taste but has no obvious effect on the growth and development of cucumber, so the NK-producing cucumber was constructed by replacing the Bt gene with NK by using CRISPR/Cas9. The pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were constructed and transformed into Agrobacterium tumefaciens EHA105, which was transformed into cucumber by floral dip method. The crude extract of NK-producing cucumber had significant thrombolytic activity in vitro. In addition, treatment with the crude extract significantly delayed thrombus tail appearance, and the thrombin time of mice was much longer than that of normal mice. The degrees of coagulation and blood viscosity as well as hemorheological properties improved significantly after crude extract treatment. These findings show that NK-producing cucumber can effectively alleviate thrombosis and improve blood biochemical parameters, providing a new direction for diet therapy against cardiovascular diseases.

Abstract

Nattokinase (NK) is effective in the prevention and treatment of cardiovascular disease. Cucumber is rich in nutrients with low sugar content and is safe for consumption. The aim of this study was to construct a therapeutic cucumber that can express NK, which can prevent and alleviate cardiovascular diseases by consumption. Because the Bitter fruit (Bt) gene contributes to bitter taste but has no obvious effect on the growth and development of cucumber, so the NK-producing cucumber was constructed by replacing the Bt gene with NK by using CRISPR/Cas9. The pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were constructed and transformed into Agrobacterium tumefaciens EHA105, which was transformed into cucumber by floral dip method. The crude extract of NK-producing cucumber had significant thrombolytic activity in vitro. In addition, treatment with the crude extract significantly delayed thrombus tail appearance, and the thrombin time of mice was much longer than that of normal mice. The degrees of coagulation and blood viscosity as well as hemorheological properties improved significantly after crude extract treatment. These findings show that NK-producing cucumber can effectively alleviate thrombosis and improve blood biochemical parameters, providing a new direction for diet therapy against cardiovascular diseases.

1 Introduction

Cardiovascular diseases (CVDs), encompassing a wide range of disorders including coronary heart disease, hypertension, cardiac hypertrophy, and heart failure, are the main cause of death and disability worldwide (Wu et al., 2020). Nattokinase (NK), a serine protease secreted by Bacillus subtilis var. natto found in the Japanese traditional food natto (Suzuki et al., 2022), possesses a variety of key beneficial effects on cardiovascular system due to its strong anti-thrombus and thrombolytic activity (Wu et al., 2019). However, the smell and viscous texture of natto are not accepted by most people, and other components in natto limit the function of NK (Nishinari et al., 2018; Liu et al., 2021).

The expression of recombinant proteins in plant systems has been promoted as a cost-effective production platform (Schillberg et al., 2019), e.g., the production of tissue plasminogen activator (Abdoli Nasab et al., 2016), LTB-VP6 (Jin et al., 2022), and HBc-based virus-like particles (Moradi Vahdat et al., 2021). Cucumber is a common economic crop plant grown around the world (Rajab et al., 2023). Because cucumber is rich in vitamins, protein, inorganic salt and low in sugar (Liu et al., 2022), it is safe for patients with CVDs. The Bt gene plays an important role in the formation of bitterness in cucumber (Shang et al., 2014). The knockout of Bt has no effect on the growth and development of cucumber and could improve fruit texture. CRISPR/Cas9-mediated gene editing has been successfully applied to bacterial, yeast, plant, and mammalian cells (Schmidt et al., 2023). This approach can be used to edit the genome at a fixed point, avoiding the destruction of endogenous genes caused by the random insertion (Wen and Zhang, 2022). Therefore, the NK gene was inserted into the cucumber genome by using CRISPR/Cas9 to stably express NK and evaluate its physiological function. This study is expected to provide a new strategy for the adjuvant prevention and treatment of CVDs.

2 Materials and methods

2.1 pZHY988-Cas9-sgRNA vector preparation

According to the sequence of the cucumber Bt gene, single-guide RNAs (sgRNAs) were designed using BioTools (http://biootools.com/). The BsaI site was added to the sgRNA and two complementary sgRNA oligos were synthesised. pZHY988-Cas9 vector was linearised by BsaI (Takara, Japan). The sgRNA was cloned into the linearised pZHY988-Cas9 vector under the action of T4 DNA ligase (Takara, Japan).

2.2 Point mutation in the NK gene

B. subtilis natto (BNCC 185324) was used to obtain the NK gene. Since the start codon of NK in the B. subtilis natto genome is GTG, it must be replaced with ATG for expression in eukaryotes. Therefore, the primers NK-M1 and NK-M2 were designed to mutate the start codon of NK.

2.3 Construction of the PX6-LHA-U6-NK-T-RHA vector

The donor gene LHA-U6-NK-T-RHA was constructed by overlap PCR and connected to the pMD-19T vector. All primers used in this study are shown in Supplementary Table 1. (Supplementary files are available on the server of the Publisher). The LHA-U6-NK-T-RHA and (Marker-free) PX6 vectors were digested by XhoI and PacI. The purified LHA-U6-NK-T-RHA and the linearised pX6 fragments were ligated by T4 DNA ligase.

2.4 Transformed pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors into Agrobacterium tumefaciens EHA105

The freezing and thawing method was used to transfer PZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors into Agrobacterium tumefaciens EHA105. The transformed cells were cultured on LB solid medium containing 50 mg mL−1 rifampicin and 50 mg mL−1 kanamycin at 28 °C for 48 h. The transformants were randomly selected for PCR identification.

2.5 Genetic transformation of cucumber by the floral dip method

Cucumber seedlings were quickly obtained by soaking the South China D9302 seeds in water (1:4, v/v) at 55 °C and stirring continuously. All female flowers were selected as genetic transformation receptors for the transformation. Female flowers 5–6 days before bloom were selected, and the A. tumefaciens EHA105 infection solution was dipped on the bud and ovary of these female flowers. It was ensured that the smear amount was large enough to drop the infection solution. The procedure was repeated again after 3 days. One day before flowering, the male flowers and infected female flowers were bagged and isolated, and the artificial pollination was conducted the next day. Cucumbers were picked and seeds were collected when the cucumbers were ripe (T0). Primers were designed according to the target sequences for PCR detection. The positive control (CT) was the PCR product obtained using pZHY988-Cas9 as template, and the negative control (WT) was the PCR product obtained using the wild-type cucumber genome as the template. The positive transgenic cucumber seeds of T0 were continuously planted according to the above method and T1 fruits were obtained for functional analyses.

2.6 Extraction and analysis of the crude extract of NK-producing cucumber

The fresh cucumber was cut into small pieces and ground into powder. The powder was mixed with PBS and centrifuged at 12,000 r.p.m. at 4 °C for 30 min. NK produced by cucumber was purified by hydrophobic chromatography and gel-filtration chromatography. The crude extract was precipitated using 60% (NH4)2SO4 and collected by centrifugation. Then, the crude extract was passed in PBS through a Sephadex G-75 column (Yuanye, Shanghai, China). Finally, the crude extract was transferred into a dialysis bag (molecular weight 8,000–14,000 Da, Yuanye, China) and put into distilled water at 4 °C for 24 h. The samples after dialysis were freeze-dried and subjected to SDS-PAGE to evaluate the purity. The activity of the crude extract was detected using the Botany Nattokinase ELISA Kit (Xinyu Biology, Shanghai, China) according to the manufacturer's instructions.

2.7 Thrombolytic effects of the crude extract of NK-producing cucumber in vitro

The thrombolytic effect of NK was measured as described previously (Choi et al., 2014). Fresh blood of a pig was allowed to clot spontaneously. The blood clot was added to the crude extract solution with different concentrations and incubated in a thermostat water bath for 5 h or 10 h at 60 r.p.m. and 37 °C. Normal saline was used as the control group. The blood clot was removed from the water bath and weighed. Dissolution of blood clot in each sample was calculated as follows:
Dissolution/%=mambma×100

Note: ma is the mass before dissolution/g; mb is the mass after dissolution/g.

2.8 Thrombus induction and NK-producing cucumber crude extract treatment in vivo

Male Kunming (KM) mice, 4 weeks of age, specific pathogen-free, were purchased from the Laboratory Animal Center of Xuzhou Medical University. After acclimatised for 1 week, the KM mice were randomly divided into four groups: (1) normal control (NC, n = 5), normal mice treated with saline, (2) negative group (NG, n = 5), intraperitoneal injection of carrageenan (50 mg kg−1) and treated with saline, (3) positive group (PG, n = 5), intraperitoneal injection of carrageenan (50 mg kg−1) and treated with nattokinase (8,000 FU kg−1), and (4) cucumber treatment group (CG, n = 5), intraperitoneal injection of carrageenan (50 mg kg−1) and treated with crude extract (8,000 IU kg−1). The time of appearance and length of the thrombus tail within 24 h were recorded. The blood was collected and the thrombin time was measured after 48 h of treatment. Fibrinogen was measured using ELISA kits (TIANGEN, Beijing, China) according to the manufacturer's instructions. The mouse tail was cut and embedded in paraffin. HE staining was used to assess the antithrombotic ability of the crude extract. The animal study protocol was approved by the Experimental Animal Ethics Committee of Xuzhou Medical University (approval number: L20210226457).

2.9 Statistical analysis

Values are presented as means ± standard deviation (SD). The data analysis was performed using SPSS program version 17. Student's t-test was used to determine differences between two groups. Multiple groups were compared by ANOVA with Dunnett's pair-wise comparisons. A value of P < 0.05 was considered statistically significant.

3 Results and discussion

3.1 pZHY988-Cas9-sgRNA vector construction

According to the cucumber Bt gene, 12 potential sgRNAs were designed. Considering the target site on the CDS of the Bt gene, the second sgRNA was selected for further analyses (Supplementary Table 2). The BsaI site was added to both ends of sgRNA by oligo synthesis. The pZHY988-Cas9 vector and sgRNA were simultaneously digested with BsaI and then connected with T4 DNA ligase. Colony PCR revealed that the length of amplification product was consistent with the expected fragment size (Fig. 1a). Sequencing results also supported the successful construction of the pZHY988-Cas9-sgRNA vector (Fig. 1b).

Fig. 1.
Fig. 1.

Construction of the pZHY988-Cas9-sgRNA vector. a. Colony PCR of the pZHY988-Cas9-sgRNA vector. Lane M: DL15000 DNA marker; Lane 1: plasmid of pZHY988-Cas9-sgRNA; Lane 2: Colony PCR products to verify pZHY988-Cas9-sgRNA construction. b. Diagram of the pZHY988-Cas9-sgRNA vector

Citation: Acta Alimentaria 52, 1; 10.1556/066.2022.00231

3.2 Point mutation in the NK gene

A single colony was randomly selected and genomic DNAs were extracted for PCR detection. The PCR product lengths were consistent with the length of the NK gene (1146 bp) (Supplementary Fig. 1a). The positive colonies were sequenced, and the sequences were consistent with the NK sequence obtained in a search against the NCBI database. The initiation codon GTG was successfully mutated to ATG (Supplementary Fig. 1b).

3.3 Construction of the pX6-LHA-U6-NK-T-RHA vector

Overlap PCR results showed that the sizes of LHA-U6 (557 bp), T-RHA (580 bp), LHA-U6-NK (1702 bp), and LHA-U6-NK-T-RHA (2282 bp) were consistent with the expected sizes (Supplementary Fig. 2). The purified LA-U6-NK-T-RA and linearised pX6 fragments were digested by XhoI and PacI and ligated by T4 DNA ligase. LHA-U6-NK-T-RHA and pX6 were successfully connected, as determined by XhoI and PacI digestion (Fig. 2a). The PCR products of full-length LA-U6-NK-T-RHA (2282 bp) were also verified by agarose gel electrophoresis (Fig. 2b).

Fig. 2.
Fig. 2.

Construction of the vector of pX6-LHA-U6-NK-T-RHA. a. Diagram of the pX6-LHA-U6-NK-T-RHA vector. b. Verification of the pX6-LHA-U6-NK-T-RHA vector. Lane M: DL15000 DNA marker; Lane 1: Recombinant vector of pX6-LHA-U6-NK-T-RHA; Lane 2: Enzyme digestion of the pX6-LHA-U6-NK-T-RHA vector by XhoI and PacI; Lane 3: Colony PCR products to verify pZHY988-Cas9-gRNA construction; Lane 4. pX6 vector

Citation: Acta Alimentaria 52, 1; 10.1556/066.2022.00231

3.4 Transformation of pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors into A. tumefaciens EHA105

Transformants showing double resistance were randomly selected for colony PCR detection. The lengths of the PCR products were nearly 400 bp and 2600 bp, consistent with the expected lengths (Supplementary Fig. 3). These results indicate that the pZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were successfully transformed into A. tumefaciens EHA105.

3.5 Genetic transformation of cucumber and genotype analysis

The cucumber cultivation and transformation processes are shown in Supplementary Fig. 4. As many female flowers were chosen as possible to improve the success rate of floral dip transformation. The fruits were picked and seeds were harvested when the cucumbers were ripe. A total of 13 cucumber fruits were harvested and 83 cucumber seeds were obtained. All harvested seeds were cultivated, and 68 seedlings survived (T0). Cucumber genomic DNA was extracted and primers were designed according to the Cas9 sequence for PCR verification. As shown in Fig. 3a (Lanes 2, 4, 5, and 6), NK-producing cucumbers were obtained.

Fig. 3.
Fig. 3.

PCR detection of T0 plants and purification of crude extract. a. Lane M: DL2500bp DNA Marker; Lane CT: PCR products of NK; Lane WT: Wild-type cucumber; Lanes 1-6: NK-producing cucumber. b. SDS-PAGE of purified crude extract. Lane M: protein Marker; Lane 1: (NH4)2SO4 precipitation of the crude extract; Lane 2: chromatography of the crude extract; Lane 3: crude extract, unpurified

Citation: Acta Alimentaria 52, 1; 10.1556/066.2022.00231

3.6 Purification of NK

NK produced by cucumber was purified by hydrophobic chromatography and gel-filtration chromatography. As shown in Fig. 3b, the crude extract yielded many bands after purification by (NH4)2SO4 precipitation (Lane 1). The bands representing impurities were reduced after purification by gel-filtration chromatography (Lane 2), and the molecular weight of the target protein was approximately 28 kDa, consistent with that of NK (Fig. 3b).

3.7 Thrombolytic activity of the crude extract of NK-producing cucumber in vitro

NK activity of the crude extract was 202.29 ± 30.70 IU mg−1, equivalent to 7.86 ± 1.10 IU g−1 (fresh cucumber). Different doses of NK were added to blood clots and dissolution was detected. As shown in Table 1, the thrombolytic effect of the crude extract increased as the concentration increased, and there was significant difference between NK groups and the control group (P < 0.001). In addition, with the extension of the reaction time, the dissolution rate for each concentration increased and was higher than that of the control group (P < 0.001). These results showed that the crude extract had significant thrombolytic activity in vitro.

Table 1.

Thrombolytic effects of the crude extract of NK-producing cucumber in vitro

GroupCrude extract (mg)ma (g)mb (g)Dissolution (%)
Control (5h)00.49 ± 0.0150.36 ± 0.03325.69 ± 7.79
Control (10h)00.52 ± 0.0330.32 ± 0.02337.17 ± 7.0
Crude extract (5h)100.51 ± 0.0420.20 ± 0.01959.51 ± 4.47***
Crude extract (5h)200.48 ± 0.0630.14 ± 0.01670.59 ± 5.58***
Crude extract (10h)100.50 ± 0.0470.19 ± 0.02262.35 ± 1.52***
Crude extract (10h)200.52 ± 0.0720.076 ± 0.01185.05 ± 3.04***

ma: the mass before dissolution/g; mb: the mass after dissolution/g; ***: P < 0.001 vs. control (5 h) or control (10 h).

3.8 Thrombolytic effects of NK-producing cucumber crude extract in vivo

Thrombin activity is an important factor causing thrombosis, and the thrombin time reflects the level of thrombin activity. In this study, the crude extract could significantly delay the appearance of the thrombus tail, and the thrombus length was significantly shorter than that of the control group (2.61 ± 0.40 cm vs. 5.22 ± 0.30 cm, P < 0.001) (Fig. 4a). After treatment with the crude extract of NK for 48 h, the thrombin time of mice was much longer than that of normal mice (12.71 ± 0.50 s vs. 7.99 ± 0.31 s, P < 0.001) (Fig. 4b). However, there was no significant difference in fibrinogen levels among the groups (Fig. 4c). Histomorphology of the thrombus tail showed that the arterial intimal became thicker, the lumen diameters increased, and the vessels were filled with blood clots in NG group compared with corresponding parameters in the NC group. Consistent with the changes in the PG group, crude extract treatment significantly reduced the degrees of coagulation and blood viscosity, inhibited intimal thickening, and improved hemorheological properties (Fig. 4d).

Fig. 4.
Fig. 4.

Effects of the crude extract on the thrombus length (a), thrombin time (b), fibrinogen levels (c), and histopathological characteristics (d) of the thrombus tail. Blood was collected to determine the thrombin time and fibrinogen concentration after treatment with the crude extract of NK for 48 h. The tails of mice were collected for a histopathological analysis after carrageenan injection for 24 h. ***: P < 0.001 vs. control. Bars correspond to 50 μm

Citation: Acta Alimentaria 52, 1; 10.1556/066.2022.00231

3.9 Discussion

Nattokinase is a thrombolytic drug with advantages over urokinase, streptokinase, and tissue plasminogen activator, including a low price, high efficiency, and lack of toxicity (Zhou et al., 2021). However, its taste limits its consumption. In this study, an adjuvant treatment scheme for CVD through diet therapy was designed by replacing the Bt gene with the NK gene by using CRISPR/Cas9, a simple, efficient, and versatile system in cucumber (Figs 12 and Supplementary Figs 1–3). Because NK can be delivered orally, cucumber as a bioreactor to produce NK can exert an adjuvant therapeutic effect by direct consumption (Wu et al., 2019). This avoids the difficulty of protein purification in plant bioreactors and the immunogenicity of plant protein injection.

“Off-target” effects are an important limitation of CRISPR/Cas9. However, these effects can be reduced by optimising gRNAs (Aquino-Jarquin, 2021). In this study, 12 sgRNAs that targeted the Bt gene of cucumber were designed, of which 9 sgRNAs had a very low probability of “off-target” effects, and one sgRNA targeting the first exon of Bt was selected in this study (Supplementary Table 2). After target gene editing using the CRISPR/Cas9 system, the homologous recombination repair system was used to replace Bt with NK. The homologous recombinant donor genes were constructed by overlap PCR, which is a PCR known to allow assembly of multiple DNA fragments into one construct (Tran et al., 2021). The extension efficiency was better when the left homologous arm was fused to the U6 promoter (LHA-U6) and the terminator was fused to the right homologous arm (T-RHA). However, the extended fusion efficiencies of larger fragments (LHA-U6-NK and LA-U6-NK-T-RHA) were reduced, and even the electrophoretic bands of PCR products were dispersed. It is possible that the overlapping region of the fusion gene was not appropriate or the fused DNA had a complex secondary structure. The length of homologous arm in eukaryotic cells greatly affects the probability of homologous recombination (Vinette and Petitclerc, 1999). However, the effect of the homologous arm length on homologous recombination efficiency in cucumber has not been reported.

4 Conclusions

In this study, 12 sgRNAs were designed using BioTools and one sgRNA was selected to connect with pZHY988-Cas9. The donor gene LHA-U6-NK-T-RHA was constructed by overlap PCR and connected with pX6. PZHY988-Cas9-sgRNA and pX6-LHA-U6-NK-T-RHA vectors were transformed into A. tumefaciens EHA105, and with the transformants the cucumbers were infected by the floral dip method. The crude extract of NK-producing cucumber has both thrombolytic and anti-coagulation functions in vivo and in vitro (Fig. 5). Therefore, NK-producing cucumber might be an ideal functional food option for the prevention and treatment of thrombosis. However, only one sgRNA for gene editing was obviously insufficient to establish the optimalised CRISPR/Cas9 system. In future studies, we will further examine the remaining sgRNAs to establish a more efficient and specific CRISPR/Cas9 gene editing system.

Fig. 5.
Fig. 5.

NK-producing cucumber was constructed using CRISPR/Cas9 and the function of its crude extract was analysed in vitro and in vivo

Citation: Acta Alimentaria 52, 1; 10.1556/066.2022.00231

Acknowledgement

This work was supported by Xuzhou Science and Technology Program (Grant number KC21273), Jiangsu Province's industry university research cooperation project (Grant number BY2022773, BY2022777), Innovation Training Program for College Students of Xuzhou University of Technology (Grant number xcx2022159).

Supplementary materials

Supplementary data to this article can be found online at https://doi.org/10.1556/066.2022.00231.

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    • Search Google Scholar
    • Export Citation
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    • Export Citation
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  • Liu, Q., Xin, D., Xi, L., Gu, T., Jia, Z., Zhang, B., and Kou, L. (2022). Novel applications of exogenous melatonin on cold stress mitigation in postharvest cucumbers. Journal of Agriculture and Food Research, 10: 100459.

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  • Liu, Y., Han, Y., Cao, L., Wang, X., and Dou, S. (2021). Analysis of main components and prospects of natto. Advances in Enzyme Research, 9(1): 19.

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  • Moradi Vahdat, M., Hemmati, F., Ghorbani, A., Rutkowska, D., Afsharifar, A., Eskandari, M.H., Rezaei, N., and Niazi, A. (2021). Hepatitis B core-based virus-like particles: a platform for vaccine development in plants. Biotechnology Reports, 29: e00605.

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  • Nishinari, K., Fang, Y., Nagano, T., Guo, S., and Wang, R. (2018). 6 - Soy as a food ingredient. In: Yada, R.Y. (Ed.), Proteins in food processing, 2nd ed. Woodhead Publishing, pp. 149186.

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  • Rajab, L., Habib, W., Gerges, E., Gazal, I., and Ahmad, M. (2023). Natural occurrence of fungal endophytes in cultivated cucumber plants in Syria, with emphasis on the entomopathogen Beauveria bassiana. Journal of Invertebrate Pathology, 196: 107868.

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  • Schillberg, S., Raven, N., Spiegel, H., Rasche, S., and Buntru, M. (2019). Critical analysis of the commercial potential of plants for the production of recombinant proteins. Frontiers in Plant Science, 10: 720730.

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  • Schmidt, T.J.N., Berarducci, B., Konstantinidou, S., and Raffa, V. (2023). CRISPR/Cas9 in the era of nanomedicine and synthetic biology. Drug Discovery Today, 28(1): 103375.

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  • Suzuki, K., Nakamura, M., Sato, N., Futamura, K., Matsunaga, K., and Yagami, A. (2022). Nattokinase, a subtilisin family serine protease, is a novel allergen contained in the traditional Japanese fermented food natto. Allergology International, https://doi.org/10.1016/j.alit.2022.11.010.

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The author instruction is available in PDF.
Please, download the file from HERE.

Senior editors

Editor(s)-in-Chief: András Salgó

Co-ordinating Editor(s) Marianna Tóth-Markus

Co-editor(s): A. Halász

       Editorial Board

  • L. Abrankó (Szent István University, Gödöllő, Hungary)
  • D. Bánáti (University of Szeged, Szeged, Hungary)
  • J. Baranyi (Institute of Food Research, Norwich, UK)
  • I. Bata-Vidács (Agro-Environmental Research Institute, National Agricultural Research and Innovation Centre, Budapest, Hungary)
  • F. Békés (FBFD PTY LTD, Sydney, NSW Australia)
  • Gy. Biró (National Institute for Food and Nutrition Science, Budapest, Hungary)
  • A. Blázovics (Semmelweis University, Budapest, Hungary)
  • F. Capozzi (University of Bologna, Bologna, Italy)
  • M. Carcea (Research Centre for Food and Nutrition, Council for Agricultural Research and Economics Rome, Italy)
  • Zs. Cserhalmi (Food Science Research Institute, National Agricultural Research and Innovation Centre, Budapest, Hungary)
  • M. Dalla Rosa (University of Bologna, Bologna, Italy)
  • I. Dalmadi (Szent István University, Budapest, Hungary)
  • K. Demnerova (University of Chemistry and Technology, Prague, Czech Republic)
  • M. Dobozi King (Texas A&M University, Texas, USA)
  • Muying Du (Southwest University in Chongqing, Chongqing, China)
  • S. N. El (Ege University, Izmir, Turkey)
  • S. B. Engelsen (University of Copenhagen, Copenhagen, Denmark)
  • E. Gelencsér (Food Science Research Institute, National Agricultural Research and Innovation Centre, Budapest, Hungary)
  • V. M. Gómez-López (Universidad Católica San Antonio de Murcia, Murcia, Spain)
  • J. Hardi (University of Osijek, Osijek, Croatia)
  • H. He (Henan Institute of Science and Technology, Xinxiang, China)
  • K. Héberger (Research Centre for Natural Sciences, ELKH, Budapest, Hungary)
  • N. Ilić (University of Novi Sad, Novi Sad, Serbia)
  • D. Knorr (Technische Universität Berlin, Berlin, Germany)
  • H. Köksel (Hacettepe University, Ankara, Turkey)
  • K. Liburdi (Tuscia University, Viterbo, Italy)
  • M. Lindhauer (Max Rubner Institute, Detmold, Germany)
  • M.-T. Liong (Universiti Sains Malaysia, Penang, Malaysia)
  • M. Manley (Stellenbosch University, Stellenbosch, South Africa)
  • M. Mézes (Szent István University, Gödöllő, Hungary)
  • Á. Németh (Budapest University of Technology and Economics, Budapest, Hungary)
  • P. Ng (Michigan State University,  Michigan, USA)
  • Q. D. Nguyen (Szent István University, Budapest, Hungary)
  • L. Nyström (ETH Zürich, Switzerland)
  • L. Perez (University of Cordoba, Cordoba, Spain)
  • V. Piironen (University of Helsinki, Finland)
  • A. Pino (University of Catania, Catania, Italy)
  • M. Rychtera (University of Chemistry and Technology, Prague, Czech Republic)
  • K. Scherf (Technical University, Munich, Germany)
  • R. Schönlechner (University of Natural Resources and Life Sciences, Vienna, Austria)
  • A. Sharma (Department of Atomic Energy, Delhi, India)
  • A. Szarka (Budapest University of Technology and Economics, Budapest, Hungary)
  • M. Szeitzné Szabó (National Food Chain Safety Office, Budapest, Hungary)
  • S. Tömösközi (Budapest University of Technology and Economics, Budapest, Hungary)
  • L. Varga (University of West Hungary, Mosonmagyaróvár, Hungary)
  • R. Venskutonis (Kaunas University of Technology, Kaunas, Lithuania)
  • B. Wróblewska (Institute of Animal Reproduction and Food Research, Polish Academy of Sciences Olsztyn, Poland)

 

Acta Alimentaria
E-mail: Acta.Alimentaria@uni-mate.hu

Indexing and Abstracting Services:

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2022  
Web of Science  
Total Cites
WoS
892
Journal Impact Factor 1.1
Rank by Impact Factor

Food Science and Technology (Q4)
Nutrition and Dietetics (Q4)

Impact Factor
without
Journal Self Cites
1.1
5 Year
Impact Factor
1
Journal Citation Indicator 0.22
Rank by Journal Citation Indicator

Food Science and Technology (Q4)
Nutrition and Dietetics (Q4)

Scimago  
Scimago
H-index
32
Scimago
Journal Rank
0.231
Scimago Quartile Score

Food Science (Q3)

Scopus  
Scopus
Cite Score
1.7
Scopus
CIte Score Rank
Food Science 225/359 (37th PCTL)
Scopus
SNIP
0.408

2021  
Web of Science  
Total Cites
WoS
856
Journal Impact Factor 1,000
Rank by Impact Factor Food Science & Technology 130/143
Nutrition & Dietetics 81/90
Impact Factor
without
Journal Self Cites
0,941
5 Year
Impact Factor
1,039
Journal Citation Indicator 0,19
Rank by Journal Citation Indicator Food Science & Technology 143/164
Nutrition & Dietetics 92/109
Scimago  
Scimago
H-index
30
Scimago
Journal Rank
0,235
Scimago Quartile Score

Food Science (Q3)

Scopus  
Scopus
Cite Score
1,4
Scopus
CIte Score Rank
Food Sciences 222/338 (Q3)
Scopus
SNIP
0,387

 

2020
 
Total Cites
768
WoS
Journal
Impact Factor
0,650
Rank by
Nutrition & Dietetics 79/89 (Q4)
Impact Factor
Food Science & Technology 130/144 (Q4)
Impact Factor
0,575
without
Journal Self Cites
5 Year
0,899
Impact Factor
Journal
0,17
Citation Indicator
 
Rank by Journal
Nutrition & Dietetics 88/103 (Q4)
Citation Indicator
Food Science & Technology 142/160 (Q4)
Citable
59
Items
Total
58
Articles
Total
1
Reviews
Scimago
28
H-index
Scimago
0,237
Journal Rank
Scimago
Food Science Q3
Quartile Score
 
Scopus
248/238=1,0
Scite Score
 
Scopus
Food Science 216/310 (Q3)
Scite Score Rank
 
Scopus
0,349
SNIP
 
Days from
100
submission
 
to acceptance
 
Days from
143
acceptance
 
to publication
 
Acceptance
16%
Rate
2019  
Total Cites
WoS
522
Impact Factor 0,458
Impact Factor
without
Journal Self Cites
0,433
5 Year
Impact Factor
0,503
Immediacy
Index
0,100
Citable
Items
60
Total
Articles
59
Total
Reviews
1
Cited
Half-Life
7,8
Citing
Half-Life
9,8
Eigenfactor
Score
0,00034
Article Influence
Score
0,077
% Articles
in
Citable Items
98,33
Normalized
Eigenfactor
0,04267
Average
IF
Percentile
7,429
Scimago
H-index
27
Scimago
Journal Rank
0,212
Scopus
Scite Score
220/247=0,9
Scopus
Scite Score Rank
Food Science 215/299 (Q3)
Scopus
SNIP
0,275
Acceptance
Rate
15%

 

Acta Alimentaria
Publication Model Hybrid
Submission Fee none
Article Processing Charge 1100 EUR/article
Printed Color Illustrations 40 EUR (or 10 000 HUF) + VAT / piece
Regional discounts on country of the funding agency World Bank Lower-middle-income economies: 50%
World Bank Low-income economies: 100%
Further Discounts Editorial Board / Advisory Board members: 50%
Corresponding authors, affiliated to an EISZ member institution subscribing to the journal package of Akadémiai Kiadó: 100%
Subscription fee 2023 Online subsscription: 776 EUR / 944 USD
Print + online subscription: 896 EUR / 1090 USD
Subscription Information Online subscribers are entitled access to all back issues published by Akadémiai Kiadó for each title for the duration of the subscription, as well as Online First content for the subscribed content.
Purchase per Title Individual articles are sold on the displayed price.

Acta Alimentaria
Language English
Size B5
Year of
Foundation
1972
Volumes
per Year
1
Issues
per Year
4
Founder Magyar Tudományos Akadémia    
Founder's
Address
H-1051 Budapest, Hungary, Széchenyi István tér 9.
Publisher Akadémiai Kiadó
Publisher's
Address
H-1117 Budapest, Hungary 1516 Budapest, PO Box 245.
Responsible
Publisher
Chief Executive Officer, Akadémiai Kiadó
ISSN 0139-3006 (Print)
ISSN 1588-2535 (Online)

 

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