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## IntechOpen Book Series Biochemistry Volume 4

Dr. Hirobumi Suzuki received his PhD in Biology from Tokyo Metropolitan University, Japan, in 1997. His research interests include the phylogeny, speciation, and embryology of the firefly as well as bioluminescence technology. He is especially interested in the genetic differentiation pattern and speciation process in relation to flashing pattern as mating behavior of some fireflies in Japan. Dr. Suzuki currently serves as a fellow at the Olympus

Corporation, a Japanese manufacturer of optics and reprography products. He is also a vice president of the Japan Fireflies Society. He and his colleagues developed a novel microscope for bioluminescence imaging that is currently being used for gene expression analysis in chronobiology, neurobiology, developmental biology, medical research, signal transduction analysis, molecular interaction analysis, and radiation biology.

**Editor of Volume 4: Hirobumi Suzuki** Olympus Corporation, Tokyo, Japan

**Book Series Editor: Miroslav Blumenberg** NYU Langone Medical Center, New York, USA

## Scope of the Series

Biochemistry, the study of chemical transformations occurring within living organisms, impacts all of life sciences, from molecular crystallography and genetics, to ecology, medicine and population biology. Biochemistry studies macromolecules proteins, nucleic acids, carbohydrates and lipids –their building blocks, structures, functions and interactions. Much of biochemistry is devoted to enzymes, proteins that catalyze chemical reactions, enzyme structures, mechanisms of action and their roles within cells. Biochemistry also studies small signaling molecules, coenzymes, inhibitors, vitamins and hormones, which play roles in the life process. Biochemical experimentation, besides coopting the methods of classical chemistry, e.g., chromatography, adopted new techniques, e.g., X-ray diffraction, electron microscopy, NMR, radioisotopes, and developed sophisticated microbial genetic tools, e.g., auxotroph mutants and their revertants, fermentation etc. More recently, biochemistry embraced the 'big data' omics systems.

Initial biochemical studies have been exclusively analytic: dissecting, purifying and examining individual components of a biological system; in exemplary words of Efraim Racker, (1913 –1991) "Don't waste clean thinking on dirty enzymes." Today however, biochemistry is becoming more agglomerative and comprehensive, setting out to integrate and describe fully a particular biological system. The 'big data' metabolomics can define the complement of small molecules, e.g., in a soil or biofilm sample; proteomics can distinguish all the proteins comprising e.g., serum; metagenomics can identify all the genes in a complex environment e.g., bovine rumen. This Biochemistry Series will address both the current research on biomolecules, and the emerging trends with great promise.

Contents

**Section 1**

**Preface III**

Bioluminescence and Applications **1**

**Chapter 1 3**

**Chapter 2 27**

**Chapter 3 43**

Bioluminescent Organisms **61**

**Chapter 4 63**

**Chapter 5 75**

**Chapter 6 87** Biofluorescence in Terrestrial Animals, with Emphasis on Fireflies: A Review

Ecological and Histological Notes on the Luminous Springtail, *Lobella* sp.

Effect of Camera Illumination on Flashing Behavior of *Pteroptyx malaccae*

*by Tadasu Sano, Yukimasa Kobayashi, Ikuko Sakai, Katsunori Ogoh* 

*by Andrew Kirkpatrick, Tingting Xu, Steven Ripp, Gary Sayler and Dan Close*

Biosensors Using Free and Immobilized Cells of Luminous Bacteria *by Anvar D. Ismailov, Leyla E. Aleskerova, Kristina A. Alenina* 

Biotechnological Advances in Luciferase Enzymes

*by Yuki Ohmuro-Matsuyama and Hiroshi Ueda*

*and Elena N. Efremenko*

*and Hirobumi Suzuki*

(Coleoptera: Lampyridae)

and Field Observation *by Ming-Luen Jeng*

*by Anchana Thancharoen and Sirima Masoh*

**Section 2**

Protein-Protein Interaction Assays Using Split-NanoLuc

(Collembola: Neanuridae), Discovered in Tokyo, Japan

## Contents


Preface

A firefly is a bioluminescent organism that produces light via a chemical reaction inside its body. But what makes a firefly glow, exactly? To answer this question, one must consider both proximate and ultimate analyses of animal behavior. A proximate analysis looks at mechanistic and ontogenic explanations, while an ultimate analysis examines evolutionary explanations for adaptation and phylogenetic history. These aspects (mechanism, ontogeny, adaption, and phylogeny) are known as Tinbergen's Four Questions. Developed by Nikolaas Tinbergen, this method was originally designed to explain animal behavior. However, this categorization is also useful for other biological traits. Hence, it is used in this book to examine the trait of bioluminescence, which is increasingly being used in biomedical applications

Section 1 on "Bioluminescence and Its Applications" contains three chapters. The first chapter examines bioengineering advances that have been applied to firefly luciferase, *Gaussia* luciferase, *Renilla* luciferase, *Ophophorus* luciferase, and bacterial luciferase. The second chapter describes a bioluminescent system called NanoLuc, which uses luciferase from the sea shrimp *Oplophorus gracilirostris*. The final chapter in this section presents a biomonitoring system of toxins in an envi-

Section 2 on "Bioluminescent Organisms" also contains three chapters. The first chapter in this section reports on the ecology and histology of the light organ of a luminous springtail discovered in Japan. The second chapter examines the effect of artificial light produced by camera illumination on the flashing behavior of synchronous fireflies in Thailand. The final chapter reviews the biofluorescence of terrestrial organisms, planarians, snails, earthworms, nematodes, velvet worms, arthropods, and vertebrates. It also describes the phenomenon of both fluorescence

I would like to express my appreciation to the chapter authors for their outstanding work, and also to Ms. Ivana Barać, Author Service Manager at IntechOpen, for effective communication and assistance during the preparation of this book.

> **Hirobumi Suzuki** Olympus Corporation,

> > Tokyo, Japan

and luminescence in fireflies using original field observations.

such as cellular assays and imaging.

ronment using photobacteria cells.
