Secretory Production of the Hericium erinaceus Laccase from Saccharomyces cerevisiae
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- Journal of Microbiology and Biotechnology
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- v.34 no.4
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- pp.930-939
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- 2024
Mushroom laccases play a crucial role in lignin depolymerization, one of the most critical challenges in lignin utilization. Importantly, laccases can utilize a wide range of substrates, such as toxicants and antibiotics. This study isolated a novel laccase, named HeLac4c, from endophytic white-rot fungi Hericium erinaceus mushrooms. The cDNAs for this enzyme were 1569 bp in length and encoded a protein of 523 amino acids, including a 20 amino-acid signal peptide. Active extracellular production of glycosylated laccases from Saccharomyces cerevisiae was successfully achieved by selecting an optimal translational fusion partner. We observed that 5 and 10 mM Ca2+, Zn2+, and K+ increased laccase activity, whereas 5 mM Fe2+ and Al3+ inhibited laccase activity. The laccase activity was inhibited by the addition of low concentrations of sodium azide and ⳑ-cysteine. The optimal pH for the 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt was 4.4. Guaiacylglycerol-β-guaiacyl ether, a lignin model compound, was polymerized by the HeLac4c enzyme. These results indicated that HeLac4c is a novel oxidase biocatalyst for the bioconversion of lignin into value-added products for environmental biotechnological applications.
(Background) Recent biotechnological breakthroughs are shedding new lights on various ethical and legal issues about human biological material. Since Rudolph Virchow, a German pathologist, had founded the medical discipline of cellular pathology, issues centering around human biological materials began to draw attention. The issues involving human biological materials were revisited with more attention along with series concerns when the human genome map was finally completed. Recently, with researches on human genes and bioengineering reaping enormous commercial values in the form of material patent, such changes require a society to reassess the present and future status of human tissue within the legal system. This in turn gave rise to a heated debate over how to protect the rights of material donors: property rule vs. no property rule. (Debate and Cases) Property rule recognizes the donors' property rights on human biological materials. Thus, donors can claim real action if there were any bleach of informed consent or a donation contract. Donors can also claim damages to the responsible party when there is an infringement of property rights. Some even uphold the concept of material patents overtaking. From the viewpoint of no property rule, human biological materials are objects separated from donors. Thus, a recipient or a third party will be held liable if there were any infringement of donor's human rights. Human biological materials should not be commercially traded and a patent based on a human biological materials research does not belong to the donor of the tissues used during the course of research. In the US, two courts, Moore v. Regents of the University of California, and Greenberg v. Miami Children's Hospital Research Institute, Inc., have already decided that research participants retain no ownership of the biological specimens they contribute to medical research. Significantly, both Moore and Greenberg cases found that the researcher had parted with all ownership rights in the tissue samples when they donated them to the institutions, even though there was no provision in the informed consent forms stating either that the participants donated their tissue or waived their rights to ownership of the tissue. These rulings were led to huge controversy over property rights on human tissues. This research supports no property rule on the ground that it can protect the human dignity and prevent humans from objectification and commercialization. Human biological materials are already parted from human bodies and should be treated differently from the engineering and researches of those materials. Donors do not retain any ownership. (Suggestions) No property rule requires a legal breakthrough in the US in terms of donors' rights protection due to the absence of punitive damages provisions. The Donor rights issue on human biological material can be addressed through prospective legislation or tax policies, price control over patent products, and wider coverage of medical insurance. (Conclusions) Amid growing awareness over commercial values of human biological materials, no property rule should be adopted in order to protect human dignity but not without revamping legal provisions. The donors' rights issue in material patents requires prospective legislation based on current uncertainties. Also should be sought are solutions in the social context and all these discussions should be based on sound medical ethics of both medical staffs and researchers.
Khori(高麗) refers to the Chaabog(reindeer) that live on lichens(蘚) on Mt. Soyon(鮮) in which pastures are the cold and dry plateau of North Eurasia. Thus, the origin region of the Khori or Koguryo that are the ancestors of the reindeer-herding pastoral nomads(馴鹿 遊牧民) can be said to be the Steppe-Taiga-Tundra pastoral areas of North Eurasia and North America. When the pastoral nomads moved on to the great mountain(大山) zone of the Jangbaek(長白) to the Baekdu(白頭) Mountains, they could have been in contact with pastoral farmers or agricultural farmers living there and they became the farmers remaining on agricultural farms. They were the Koryo people, the ancestors of Korea. Staying in one place, they gradually forgot the origin of their reindeer-herding pastoral nomadic history in the Northwest area of Mt. Soyon, the small mountain(小山) zone of the Steppe-Taiga-Tundra pastoral areas. In other words, they lost their identity as reindeer-herding pastoral nomads when they entered the agricultural area after leaving the pastoral area. However, since their basic genes had already formed when they lived on the cold and dry plateau of North Eurasia, it is possible to study their pastoral nomadic history focusing on 'the minority living in the broad area(廣域少數)', by utilizing highly advanced biotechnological science and focusing on genes and information technology innovation, and removing various past hindrances in research. Therefore, it is not so difficult to restore the reindeerherding pastoral nomadic history of the Koguryo(高句麗) people and secure their pastoral nomadic identity, of which the first steps have already been taken into their historical stages. The Eurasian continent and the Korean peninsula, especially the cold and dry plateau of North Eurasia and the Korean peninsula have been closely related to each other ecologically and historically. They can never be a separate space at all. The Eurasian continent lies horizontally east to west and thus, the continent forms an isothermal zone. Also, since the time of producing their own foods, it was relatively easy for people with their technology to move to other places owing to the pastoral nomadic characteristic of mobility. Unlike the Chungyen(中原) region, western Asia and the regions covering the Siberia-Manchu-Korean peninsula where food production revolution was first made were connected to the Mongolian lichens route(蘚苔之路: Ni, ukinii jam) and steppe roads. Although the ecological conditions of nature have changed a bit throughout a long history, it was natural for the many tribes in North Asia living on the largest Steppe-Taiga-Tundra area in the world to have believed 'the legends related to animals in relation to their founders and ancestors(獸祖傳說)'. Assuming that Siberian tigers and the tigers living on Mt. Baekdu were connected ecologically and genetically because of the ecological characteristics of the animals, and their migration from plateau to plateau, we would suspect that the Chosun(朝鮮) tribe living on Mt. Baekdu were ethnically and culturally more closely connected to the farther removed Ural-Altai tribes that lived on the cold and dry plateau region than to the Han(i14;) tribe who lived in Chungyen(中原) that was close to Mt. Baekdu. More evidence is the structure of the Korean language which has the form of 'Subject + Object + Verb', which is assumed to have originated from the speedy lifestyle of the reindeer-herding pastoral nomads. The structure is quite different from that of the Han(漢) language, which is based on agricultural life. Also, it is natural for reindeer riding reindeerherding pastoral nomads or horse-riding sheep-herding pastoral nomads(騎馬, 羊遊牧民) to have held military and political power over the region and eventually to have established an ancient pastoral nomadic empire in the process of their conquest of agricultural regions. The stages for founding global empires in the history of mankind maybe largely divided into two, in terms of ecological conditions and occupations. They are the steppes and the oceans. Of course, the steppe-based empires were established based on the skills to deal with horses and the ability to shoot arrows while riding horses, along with the use of iron ware in the 8th century BC. The steppe-based empires became the foundation for an oceanic empire, which could have been established by the use of warships and warship guns since the 15th Century. Based on those facts, we know that Chosun, Puyo(夫餘), and Koguryo are the products of a developmental process of pastoral nomadic empires on the steppes. Maybe we can easily find the pastoral nomadic identity of the Koguryo more than we expected when we trace the origins and history of the Korean tribe living in the pastures located in the northwest area of Mt. Jangbaek by focusing on pastoral nomadic mobility and organization just as we have investigated the historic origins of Anglo-Saxons in America by focusing on the times before the 15th Century. In the process, we should keep in mind that English culture originated from the Industrial Revolution and was directly delivered to the American continent, although America was far from England and was not an intermediate point on long sojourns either. Further, American culture came back to England in a more advanced form later. The most important thing currently to be resolved is to cause Koreans to look back on their own history in a freer way of thinking and with diverse, profound, and sharp insight, taking away the old and existing conventional recognition that is entangled with complicated interests with Korean people and other countries. The meanings of Chosun, Khori, and Solongos have been interpreted arbitrarily without any historic evidence by the scholars who followed conventional tradition of fixed-minded aristocrats in an agricultural society. If the Siberian cultural properties of the stone age, the earthenware age, the bronze age, and the iron age are analyzed in such a way, archaeological discovery will never be able to contribute to the restoration of the Koguryo's pastoral nomadic identity. One should transcend the errors that tend to interpret the cultural properties discovered in the pastoral nomadic regions as not being differentiated from those of agricultural regions and just interpret them altogether from the agricultural point of view. A more careful intention is required in the interpretation of cultural properties of ancient Korean empires that seem to have been formed due to mutual interactions of pastoral nomadic and agricultural cultures. Also, it is required that the conventional recognition chain of 'reverse-genes' be severed, which has placed more weight on agricultural properties than pastoral nomadic ones, since their settlement on agricultural farms was made after the establishment of their ancient pastoral nomadic empires. There is no reason at all to place priority on stoneware, earthenware, bronze ware, and iron ware than on wooden ware(木器) and other ware which were made of animal skins(皮器), bones and horns(骨角器), in analyzing the history in the regions of reindeer or sheep pastures. Reading ancient Korean history from the perspective of pastoral nomadic history, one feels strongly the instinctive emotions to return to the natural 'mother place'. The reindeer-herding pastoral nomadic identity of the Koguryo people that has been accumulated in volumes in their genes and hidden deep inside and have interacted organically could be reborn with Burqanism(Burqan refers to 不咸 in Chinese), which was their religion by birth and symbolized as the red willow(紅柳=不咸). The mother place of the Koguryo's people is the endless vast green pastures of North Eurasia and North America, where we anticipated the development of Korean livestock farming following the inherent properties in the genes of the reindeer-herding pastoral nomads with Korean ancestors. We anticipate that the place would be the core resource that could contribute to the development of life of living creatures following the inherent properties of their genes and biotechnological factors. In other words, biotechnology used for a search for clues on the well-being of humans could be the fruit brought by Burqanism of the Koguryo people and the fruit of the globalization of Korean livestock farming. It is the Chosun farmer in China come from the vast nomadic reindeer pastures of North Eurasia that resolved the food problem of a billion Chinese people with lowland paddy rice seeds (水稻) by transforming Heilongjiang Province(黑龍江省) into an oceanic lowland paddy rice field(水田). Even Mao Tse-tung(毛擇東) could not resolve the food problem by his revolution campaigns for tens of years. Today is the very time that requires the development of special livestock farming following the inherent properties of the ancient Korean reindeer-herding pastoral nomads that respected the dignity of life on the cold and dry plateau of North Eurasia and the America continent. I suggest that research should be started from the pastures of the Dariganga Steppe in East Mongolia that was the homeland of Hanwoo(韓牛) and the central horse-herding steppe place(牧馬場) of Chingis Khan's Mongolia. The Dariganga Steppe is awash with an affluent natural environment for pastoral nomadic living however, the quality of life of the pastoral nomads there is still low. I suggest we Koreans, the descendents of the Koguryo, should take our first steps for our livestock farming business project and develop the Northern nomadic pastures, here at the pastures of the Dariganga Steppe, which is the Mongolian core place of state-of-the-art technology for military weapons.