Purification and characterization of a novel milk-clotting metalloproteinase from Paenibacillus spp. BD3526

Feng, Hao; Wang, Qinbo; Hong, Qing; Liu, Peiyi; Wu, Zhengjun; Liu, Zhenmin; Zhang, Hao; Chen, Wei

doi:10.1016/j.ijbiomac.2016.01.028

Cited by 23 publications

(24 citation statements)

References 39 publications

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“…Milk-clotting peptidases belonging to albizia and ginger cleaved k-casein in other sites, such as Lys 116 -Thr 117 , Ala 90 -Glu 91 and His 102 -Leu 103 , respectively (Egito et al, 2007;Huang, Chen, Luo, Guo, & Ren, 2011). It has also been described that these slight differences in cleavage sites do not affect milk-clotting activity and taste of the cheeses (Hang et al, 2016).…”

Section: Latex Peptidasesmentioning

confidence: 99%

Insights into milk-clotting activity of latex peptidases from Calotropis procera and Cryptostegia grandiflora

Freitas

Leite

Oliveira

et al. 2016

Food Research International

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Latex fractions from Calotropis procera, Cryptostegia grandiflora, Plumeria rubra, and Himatanthus drasticus were assayed in order to prospect for new plant peptidases with milk-clotting activities, for use as rennet alternatives. Only C. procera and C. grandiflora latex fractions exhibited proteolytic and milk-clotting activities, which were not affected by high concentrations of NaCl and CaCl. However, pre-incubation of both samples at 75°C for 10min eliminated completely their activities. Both proteolytic fractions were able to hydrolyze k-casein and to produce peptides of 16kDa, a similar SDS-PAGE profile to commercial chymosin. RP-HPLC and mass spectrometry analyses of the k-casein peptides showed that the peptidases from C. procera or C. grandiflora hydrolyzed k-casein similar to commercial chymosin. The cheeses made with both latex peptidases exhibited yields, dry masses, and soluble proteins similar to cheeses prepared with commercial chymosin. In conclusion, C. procera and C. grandiflora latex peptidases with the ability to coagulate milk can be used as alternatives to commercial animal chymosin in the cheese manufacturing process.

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Section: Latex Peptidasesmentioning

confidence: 99%

Insights into milk-clotting activity of latex peptidases from Calotropis procera and Cryptostegia grandiflora

Freitas

Leite

Oliveira

et al. 2016

Food Research International

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“…The SDS‐PAGE results also indicated a broad specific activity of the microbial rennet from the rice wine on milk caseins, as reported for other microbial proteases, such as an extracellular metalloprotease from the edible mushroom Termitomyces clypeatus (Majumder, Banik, & Khowala, ) and the milk‐clotting enzyme from Paenibacillus spp . BD3526 (Hang et al, ). The proteolytic pattern of milk proteins by the microbial rennet of this study suggested a possible effect of the enzyme on the cheese quality as a result of its proteolytic activity.…”

Section: Resultsmentioning

confidence: 99%

Change of proteolysis and sensory profile during ripening of Cheddar‐style cheese as influenced by a microbial rennet from rice wine

Zhao

Zheng

Zhang

et al. 2019

Food Science & Nutrition

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To test the potential of a novel microbial rennet isolated from traditional fermented rice wine for cheese making, Cheddar‐style cheese made with this enzyme was studied for changes in composition, proteolysis, and sensory profile during 90 days of ripening in comparison with a control cheese made with a commercial rennet. The initial proteolysis assay of the microbial rennet on milk proteins indicated a notable increase in the hydrolysis of casein components (α‐, β‐, and κ‐caseins) but no effect on whey proteins upon increasing the concentration of the enzyme. Correspondingly, compared to cheese made with commercial rennet, the use of the microbial rennet in Cheddar‐style cheese resulted in significantly higher primary and secondary proteolysis in the later stages of ripening (60–90 days ripening) and thus a softer texture and the formation of more volatile compounds and free amino acids (FAAs) despite its lower moisture content (41.7%, w/w). Though the cheese made with the microbial rennet was found to contain bitter‐taste FAAs (1,000 mg/100 g), the combined effect of other‐taste FAAs, including sweet (231 mg/100 g), umami (225 mg/100 g), and tasteless (361 mg/100 g) FAAs, in the cheese attenuated the bitter taste of the cheese. This analysis was in accordance with the sensory evaluation, which showed no significantly different sensory scoring between the cheeses made with the microbial and commercial rennets. The present study demonstrated a novel approach to evaluate the bitter taste of ripened cheese. The results of this study suggest the potential of the microbial rennet from rice wine to serve as a new source of milk‐clotting agents in cheese making.

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“…The active fractions were eluted in 2 M NaCl, which was similar to the purification of MCE from Paenibacillus spp. BD3526 using a DEAE Sepharose Fast Flow chromatography column (Hang et al, 2016a;2016b). The MCE from this step gave a single protein band on SDS-PAGE ( Fig.…”

Section: Purification Of the Mce From B Methanolicus Lb-1mentioning

confidence: 99%

“…Many bacteria, especially the species belonging to Bacillus such as B. subtilis (Wu et al, 2013), B. amyloliquefaciens (An et al, 2014) and Paenibacillus spp. (Hang et al, 2016a;2016b), have been reported to produce extra-cellular MCEs. Production of bacterial MCEs by submerged fermentation (SMF) was efficient with a high yield of MCE (Ding et al, 2012), and it was simple to operate compared to solid state fermentation (SSF) by fungus (Thakur et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

“…Unfortunately, despite many reports on the production, purification and characterization of various bacterial MCEs, few of them met the requirement of an ideal MCE (Sato et al, 2004;Zhao et al, 2015). Production of bacterial MCEs was influenced by cultivation temperature, filled volume of growth medium, cultivation time and rotation speed in shaking-flask cultivation (Hang et al, 2016a;2016b). Consequently, the ideal MCE could be produced by screening new bacterial producing strains and optimizing fermentation conditions.…”

Section: Introductionmentioning

confidence: 99%

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Production, purification and characterization of a milk clotting enzyme from Bacillus methanolicus LB-1

Zheng

Zhao

et al. 2019

Food Sci Biotechnol

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Bacillus methanolicus LB-1 isolated from traditional rice wine was found to produce a milk clotting enzyme (MCE), and its fermentation conditions were optimized using response surface methodology. Then the MCE was produced by ethanol precipitation, and further chromatography separation resulted in a 10.46-fold purification with a 59.28% recovery. The MCA (milk clotting activity) of the purified MCE reached 597,310 ± 0.13 SU/ g. The optimal temperature of the MCE was determined to be 50°C and it was stable in the low temperature range of 40-45°C. The MCE had an optimum pH of 6.5, and it was stable under neutral conditions. Calcium chloride at the concentration of 25 mM was found to be the most effective stimulus. The MCE was identified by LC-MS to be a putative protein (ID I3EB99) containing 759 amino acids with a molecular weight of 80.37 kDa and a pI of 9.23.

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Purification and characterization of a novel milk-clotting metalloproteinase from Paenibacillus spp. BD3526

Cited by 23 publications

References 39 publications

Insights into milk-clotting activity of latex peptidases from Calotropis procera and Cryptostegia grandiflora

Insights into milk-clotting activity of latex peptidases from Calotropis procera and Cryptostegia grandiflora

Change of proteolysis and sensory profile during ripening of Cheddar‐style cheese as influenced by a microbial rennet from rice wine

Production, purification and characterization of a milk clotting enzyme from Bacillus methanolicus LB-1

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