Papain-like cysteine proteases in Carica papaya: lineage-specific gene duplication and expansion

Liu, Juan; Sharma, Anupma; Niewiara, Marie Jamille; Singh, Ratnesh; Ming, Ray; Yu, Qingyi

doi:10.1186/s12864-017-4394-y

Cited by 32 publications

(30 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Most of the enzymes will disappear with the ripening of fruits (Payasi, Mishra, Chaves, & Singh, ). For example, protease activities of figs and papayas are very high in the green fruits but become undetectable in softening ones (Liu et al, ; Zare et al, ). Pineapple plants produce bromelain in a different mode to other plants, as almost similar amounts of bromelain are present at the early and at the late stages of pineapple fruit development (Rajni, ).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Fractionation and activity profiling of fruit bromelain from pineapples of Phuket variety growing in Thailand

et al. 2019

View full text Add to dashboard Cite

Bromelain is a type of protease found in both fruits and stems of pineapples. Stem bromelain has been extensively studied and is commercially available for applications in various industries. In contrast, studies of fruit bromelain are quite limited since most of pineapples have been consumed freshly, canned or juiced. Nowadays, the consumption of canned fruits, including canned pineapples has decreased greatly. Fruit bromelain could be a new growth point for pineapple industry. In this study, fruit bromelain was extracted from the pineapple juice of Phuket variety and some of its properties were studied. The enzyme was purified by precipitation using ammonium sulfate fractionation followed by ion‐exchange and gel filtration chromatography. Consequently, the protease purification level was increased by 95.2 fold. The final specific activity was getting to 448,590 U/mg on average, dominated by cysteine proteases, with optimal activity at 45°C and pH ranging from 6 to 8. The study facilitates the molecular and application research of fruit bromelain. Practical applications The research has been carried out at Funong Food Technology Co., Ltd., Guangdong, China, which produces primarily pineapple chunks and juice. As plenty of by‐products, like peels and cores of pineapples, are produced, the techniques are employed to extract bromelain from the by‐products. The techniques reported in this work are not new or advanced, however, they are applicable during the manufacturing process and the related equipment is easy to set up and operate. What's more, the practical application of the techniques is cost‐effective for the manufactories. Take Funong Food Technology Co., Ltd. as an example, they was using 80% saturation ammonium sulfate to precipitate protein from pineapple juice and obtained a bromelain with activity of approximately 8,000 U/mg and yield of 1.7 kg per ton of juice. With the application of the techniques reported in this work, bromelain was first extracted by ammonium sulfate gradient precipitation, and then purified through ion‐exchange and gel filtration chromatography. Each step of precipitation and purification generates a different level of activity and recovery of bromelain, ranging from around 2,506 to 448,590 U/mg, which allows for the production of bromelain according to the requirement of the market and brings more profits.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Most of the enzymes will disappear with the ripening of fruits (Payasi, Mishra, Chaves, & Singh, 2009). For example, protease activities of figs and papayas are very high in the green fruits but become undetectable in softening ones (Liu et al, 2018;Zare et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Fractionation and activity profiling of fruit bromelain from pineapples of Phuket variety growing in Thailand

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Thus, serine protease was the main protease class expressed in the fifth instar. High cysteine protease activities have already been detected in studies involving young larvae of Lepidoptera (Liu et al, ; Senthil‐Nathan, ) as well as the control of the gene expression of these enzymes, showing that there is an increase in the synthesis of serine proteases rather than the decrease of cysteine proteases in Tribolium castaneum (Lomate et al, ).…”

Section: Resultsmentioning

confidence: 95%

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars

Junior

Vital

Barros

et al. 2019

Arch Insect Biochem Physiol

View full text Add to dashboard Cite

Soybean is one of most consumed and produced grains in the world, and Anticarsia gemmatalis is a pest that causes great damage to this crop due to severe defoliation during its larval phase. Plants have mechanisms that lead to the inhibition of proteases in the intestine of these herbivores, hampering their development. Understanding this complex protease inhibitor is important for pest control. The objective of this study was to evaluate the enzymatic profiles of the intestinal proteases of the soybean caterpillar at different instars. For this, the proteolytic profile of the gut in the third, fourth, and fifth instars were analyzed.Irreversible inhibitors of proteases were separately incubated with A. gemmatalis enzyme extracts at the third, fourth, and fifth instar to assess the contribution of these proteases to total proteolytic activity. The enzymatic extracts were also evaluated with specific substrates to confirm changes in the specific activities of trypsin-like, chymotrypsin-like, and cysteine proteases at different instars. The results showed that the protease profile of A.gemmatalis gut changes throughout its larval development.The activity of cysteine proteases was more intense in the first instar. On the contrary, the serine proteases The protease profile of the gut of A. gemmatalis changes throughout larval development. In the third instar, cysteine protease activity is predominant over others. In the fourth and fifth instars, proteolytic activity is mainly due to serine proteases. This difference in activity in the different instars of A. gemmatalis suggests that the expression of the genes coding for proteases also changes throughout the development of the insect. Using serine PIs is a good strategy for controlling insect pests, as these are the main proteolytic enzymes of Lepidoptera. However, the presence of other important protease subclasses in insect digestibility suggests that a combination of serine and cysteine PIs may be promising, aiming to compromise larval development at different instars.

show abstract

“…In this group, 181 members were identified. With A. thaliana , 31 proteases were reported [52], 33 in rice [17], 43 in rubber [53] and 33 in papaya [43]. Subfamilies of PLCPs (already described for A. thaliana [52] and rice [17]) were also identified.…”

Section: Discussionmentioning

confidence: 99%

“…Liquid chromatography-mass spectrometry (LC-MS) is a powerful technique, which has been successfully used to study the properties of different wheat cultivars [2,3,39], plant development [40,41], and wheat responses to biotic [35,36] and abiotic stresses [37]. An LC-MS approach has been adopted in this study for characterization of all wheat proteases that were identified using Pfam identificators attributed to all the proteases with the use of profile hidden Markov model (HMMER) algorithm [42] that was previously successfully used for identification of proteins [43]. We performed classification of identified proteases.…”

Section: Introductionmentioning

confidence: 99%

Proteomics Analysis Reveals That Caspase-Like and Metacaspase-Like Activities Are Dispensable for Activation of Proteases Involved in Early Response to Biotic Stress in Triticum aestivum L.

Balakireva

Deviatkin

Zgoda

et al. 2018

IJMS

View full text Add to dashboard Cite

Plants, including Triticum aestivum L., are constantly attacked by various pathogens which induce immune responses. Immune processes in plants are tightly regulated by proteases from different families within their degradome. In this study, a wheat degradome was characterized. Using profile hidden Markov model (HMMer) algorithm and Pfam database, comprehensive analysis of the T. aestivum genome revealed a large number of proteases (1544 in total) belonging to the five major protease families: serine, cysteine, threonine, aspartic, and metallo-proteases. Mass-spectrometry analysis revealed a 30% difference between degradomes of distinct wheat cultivars (Khakasskaya and Darya), and infection by biotrophic (Puccinia recondita Rob. ex Desm f. sp. tritici) or necrotrophic (Stagonospora nodorum) pathogens induced drastic changes in the presence of proteolytic enzymes. This study shows that an early immune response to biotic stress is associated with the same core of proteases from the C1, C48, C65, M24, M41, S10, S9, S8, and A1 families. Further liquid chromatography-mass spectrometry (LC-MS) analysis of the detected protease-derived peptides revealed that infection by both pathogens enhances overall proteolytic activity in wheat cells and leads to activation of proteolytic cascades. Moreover, sites of proteolysis were identified within the proteases, which probably represent targets of autocatalytic activation, or hydrolysis by another protease within the proteolytic cascades. Although predicted substrates of metacaspase-like and caspase-like proteases were similar in biotrophic and necrotrophic infections, proteolytic activation of proteases was not found to be associated with metacaspase-like and caspase-like activities. These findings indicate that the response of T. aestivum to biotic stress is regulated by unique mechanisms.

show abstract

Papain-like cysteine proteases in Carica papaya: lineage-specific gene duplication and expansion

Cited by 32 publications

References 59 publications

Fractionation and activity profiling of fruit bromelain from pineapples of Phuket variety growing in Thailand

Fractionation and activity profiling of fruit bromelain from pineapples of Phuket variety growing in Thailand

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars

Proteomics Analysis Reveals That Caspase-Like and Metacaspase-Like Activities Are Dispensable for Activation of Proteases Involved in Early Response to Biotic Stress in Triticum aestivum L.

Contact Info

Product

Resources

About