Biotechnological production and practical application of L-asparaginase enzyme

Figures & data

Figure 1. General mechanism of L-asparaginase action.

Table 1. L-asparaginase producing micro-organisms.

L-asparaginase sources	Name of species	References
Bacteria	Pseudomonas aeruginosa 50071, Pyrococcus furiosus, E. chrysanthemi, Helicobacter pylori, Mycobacterium phlei, Pseudomonas Stutzeri MB-405, Thermus thermophilus, Tetrahymena pyriformis, Bacillus polymyxa, Bacillus circulans, Corynebacterium glutamicum, Nocardia levis MK-VL_113, Lactobacillus crispatus strain CLS01, Enterobacter aerogenes, Azotobacter vinelandii, Listeria monocytogens, Lactobacillus amylovorous, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus fermentum, Lactobacillus rhamnnosus, Bacillus cereus, Bacillus subtilis, Bacillus megaterium, Bacillus pasteurii, Bacillus amyloliquefacciens, Cellulomonas fimi, Cellulomonas uda, Klebsiella pneumoniae, Salmonella paratyphi A-1, Salmonella enteritidis, Staphylococcus aureus, Shigella somei, Shigella flexneri, Proteus vulgaris, Pseudomonas putida, Shewanella putrifaciens	El-Bessoumy, Sarhan, and Mansour (2004), Vimal and Kumar (2017)
Fungi	Fusarium equisetti, Aspergillus flavus (KUFS 20), Aspergillus terreus, Aspergillus nidulans, Penicillum sp., A. tamari, Emericella nidulans, Fusarium oxysporum, Aspergillus niger,, Aspergillus albus, Aspergillus sclerotiorum, Penicillum notatum, Penicillum frequentans, Penicillum citrinum, Penicillum purpurogenum, Penicillum lilacinum, Serpula lacrymans, Volvariella volvacea, Agaricus bisporous, Aureobasidium bisporous	Hosamani and Kaliwal (2011), Vimal and Kumar (2017)
Actinomycetes	Streptomyces Karnatakenesis, S. venezuelae, Thermoactinomyces vulgaris, Streptomyces gulbargensis, Streptomyces tendae TK-VL-333	Carta de-Angeli et al. (1970)
Plant	Tamarindus indica, Capsicum annum L., Withania somnifera, Lupinus angustifolius, Pisum sativum, Pisum arvense, Vicia faba, Phaseolus multiflorus, Lipinus polyphyllus, L. albus var. Buttercup, L. mutabilis var. Tabor, Zea mays var. Fronica, Hordeumvulgare var. Julia, Pinus pinaster, Pinus radiate, Lupin angustiplius, Arabidopsis thaliana, Lupinus luteus, Amaranthus polygonoides	Bano and Sivaramakrishnan (1980)
Yeast	Candida utilis, Pichia polymorpha, Saccharomyces cerevisiae, Candida glabrata, Candida krusei, Candida dublinensis, Candida parapsilosis, Candida kefyr	Kil, Kim, and Park (1995), Vimal and Kumar (2017)
Algae	Chlamydomonas sp., Chlorella vulgaris	Paul and Cooksey (1981)
Animal	Rattus norvegicus, Homosapiens	Wriston and Yellin (1973)

Table 2. L-asparaginase production by submerged fermentation at various operating condition.

Micro-organism	Medium	Temp (°C)	pH	Incubation period (hrs)	Rpm	Activity	References
Spirulina maxima	Modified Zarrouk medium	25 ± 3	9.5	432	–	51.28 IU/L	Abd El Baky and El Baroty (2016)
Enterobacter aerogenes MTCC111	Trisodium citrate (.75%)	33	6	40		18.35 IU/ml	Erva, Goswami, and Suman (2016)
	Ammonium chloride(.15%)
Pectobacterium carotovorum MTCC 1428	Glucose (30 g/L)	30	8.5	Fed-batch mode		38.8 U/mL	Kumar et al. (2017)
	L-asparagine (30 g/L)
Penicillium digitatum	Czapek–Dox medium	37	–	96	–	363.80 IU/ml	Shrivastava et al. (2012)
Nocardia levis MK-VL 113	Asparagine-glycerol salts (ISP-5) broth	30	7	72	–	5.06 ± .002 U/mg of cell dry wt.	Kavitha and Vijayalakshmi (2012)
Aspergillus terreus MTCC 1782	Czapek–Dox Medium + L-asparagine 1%, yeast extract 1%, peptone .6%, glucose .4%	35	6	72	160	24.10 IU/ml	Baskar and Sahadevan (2011)
Emericella nidulans	Czapek–Dox Medium	30	6	48	–	1.1 IU	Jayaramu, Hemalatha, Rajeshwari, Siddalingeshwara, and Sunil (2010)
Streptomyces Albidoflavus	Asparagine-maltose-yeast extract-salts broth	35	7.5	72	–	7.51 IU/mg of cell dry weight	Narayana, Kumar, and Vijayalakshmi (2008)
E.coli K-12	Lactose; 10 g/l, Tryptone; 10 g/l, yeast extract; 5 g/l, L-asparagine; 2 g/l and CaCl2; 15 g/l	37	6.5		200	3.82 ± .1 IU/ml	Kumar, Punia, and Verma (2012)
Streptomyces sp (TA22)	Sucrose-potassium nitrate-1% asparagine	28	7	120	120	8.87 IU/mg	Priya et al. (2011)
Bacillus sp. (DKMBT10)	KH2PO4 2.0, L-asparagine 6.0, MgSO4.7H2O 1.0, CaCl2.2H2O 1.0 and glucose/maltose 3.0)	37	7	24	200	.1 U/mg	Moorthy, Ramalingam, Sumantha, and Shankaranaya (2010)
Pseudomonas fluorescens	Glucose, beef extract, L-Asparagine, salt solution	37	8	48	–	168.4 IU/ml	Prema et al. (2013)

Note: IU-International unit.

Table 3. L-asparaginase production by solid state fermentation at various operating condition.

Micro-organism	Substrate		Moisture (%)	Temp (°C)	pH	Incubation period (hrs)	Activity	References
Serratia marcescens (NCIM 2919)	Coconut oil cake(OFAT)	6 g	40	35	6	24	3.87 U/gds	Ghosh et al. (2013)
Serratia marcescens(NCIM 2919)	COC (by response surface method)	7.6 g	50	35.5	7.4	24	5.86 U/gds	Ghosh et al. (2013)
Serratia marcescens-NCIM 2919	Citrus limetta pulp	5 g	60	28	7.5	48	83.16 U/g	Kumar, Thangabalan, and M (2011)
Serratia marcescens SB08	Rice bran	10 g	50	30	7	36	79.84U/g	Venil and Lakshmanaperumalsamy (2013)
Pseudomonas aeruginosa 50071	Soya bean meal	10 g	50	37	7.4	96	1900 IU/mg	El-Bessoumy et al. (2004)
Aspergillus niger	Soya bean (30 min of cooking time)		70	30	6.5	96	(40.9 +/−3.35 U/gds	Mishra (2006)
Aspergillus terreus MTCC 1782	Pomegranate	5 g	75	30		120	253 U/gds	Nair, Kumar, Devi, and Balakrishnan (2013)
Aspergillus terreus MTCC 1782	Wheat bran	5 g	75	30		72	110 U/gds	Nair et al. (2013)
Aspergillus terreus MTCC 1782	M9 medium moistened coconut oil cake	5 g	75	30		72	85 U/gds	Nair et al. (2013)
Aspergillus terreus MTCC 1782	Bajra seed flour	5 g	70	30	8	96	273.3 U/gds	Varalakshmi and Raju (2013)
Aspergillus terreus MTCC 1782	Corn cob	5 g	70	30	8	96	55.66 U/gds
Aspergillus terreus KLS2	Carob pod	20 g	65	35	4.5	168	5.63 IU	Jayaramu et al. (2010)
Fusarium oxysporum	Wheat bran	5 g	60	30	7	120	8.14 IU	Pallem, Nagarjun, and Srikanth (2011)
Aspergillus flavus (KUFS20)	Orange peel	20 g	40	35	6	96	339.16 U/g	Soniyamby et al. (2011)
Aspergillus terreus MTCC 1782	Sesame oil cake(SOC)	5 g	40	30		96	68.49 U/gds	Suresh and Raju (2013)
Aspergillus terreus MTCC 1782	Black gram husk(BH)	5 g	40	30		96	15.95 U/gds	Suresh and Raju (2013)
Aspergillus terreus MTCC 1782	(SOC + BH) (7:3)	5 g	40	30		96	74.21 U/gds	Suresh and Raju (2013)
Aspergillus terreus MTCC 1782	(SOC + BH) (7:3)	5 g	60	32	7	120	163.34 U/gds	Suresh and Raju (2013)
Serratia marcescens-NCIM 2919	Sesame oil cake	10 g	68.64	37		96	110.795 U/gds	Vuddaraju, Nikku, Chaduvula, Dasari, and Donthireddy (2010)

Notes: IU – International Unit, U/gds – Units/ gram dry solids, U/g – Unit/ gram.

Figure 2. Various potential applications of L-asparaginase.

Figure 3. Group A streptococcus proliferation in presence of high level of L-asparagine. Infection could be checked by introducing L-asparaginase that deaminates the L-asparagine needed for pathogen growth and development.

Table 4. Reduction in acrylamide content in different food items after using Acrylaway.

Food items	Acrylamide reduction (%)
Biscuits and cookies	50–90
Crisp bread and toasted bread	50–90
Crackers	75–85
Snacks	75–90
Coffee	40–70
French fries	35–50
Breakfast cereals	55–70

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